scholarly journals Insecticide resistance selection and reversal in two strains of Aedes aegypti

2020 ◽  
Vol 5 ◽  
pp. 183
Author(s):  
Jonathan Thornton ◽  
Bruno Gomes ◽  
Constância Ayres ◽  
Lisa Reimer
Keyword(s):  
Aedes Aegypti ◽  
Insecticide Resistance ◽  
Vector Competence ◽  
Resistance Mechanisms ◽  
Target Site ◽  
Metabolic Resistance ◽  
Resistance Selection ◽  
Similar Phenotype ◽  
Target Site Resistance ◽  
Homozygous Resistant

Background: Laboratory reared mosquito colonies are essential tools to understand insecticide action. However, they differ considerably from wild populations and from each other depending on their origin and rearing conditions, which makes studying the effects of specific resistance mechanisms difficult. This paper describes our methods for establishing multiple resistant strains of Aedes aegypti from two colonies as a new resource for further research on metabolic and target site resistance. Methods: Two resistant colonies of Ae. aegypti, from Cayman and Recife, were selected through 10 generations of exposure to insecticides including permethrin, malathion and temephos, to yield eight strains with different profiles of resistance due to either target site or metabolic resistance. Resistance ratios for each insecticide were calculated for the selected and unselected strains. The frequency of kdr alleles in the Cayman strains was determined using TaqMan assays. A comparative gene expression analysis among Recife strains was conducted using qPCR in larvae (CCae3A, CYP6N12, CYP6F3, CYP9M9) and adults (CCae3A, CYP6N12, CYP6BB2, CYP9J28a). Results: In the selected strain of Cayman, mortality against permethrin reduced almost to 0% and kdr became fixated by 5 generations. A similar phenotype was seen in the unselected homozygous resistant colony, whilst mortality in the susceptible homozygous colony rose to 82.9%. The Recife strains showed different responses between exposure to adulticide and larvicide, with detoxification genes in the temephos selected strain staying similar to the baseline, but a reduction in detoxification genes displayed in the other strains. Conclusions: These selected strains, with a range of insecticide resistance phenotypes and genotypes, will support further research on the effects of target-site and/or metabolic resistance mechanisms on various life-history traits, behaviours and vector competence of this important arbovirus vector.

scholarly journals Insecticide resistance selection and reversal in two strains of Aedes aegypti

2020 ◽  
Vol 5 ◽  
pp. 183
Author(s):  
Jonathan Thornton ◽  
Bruno Gomes ◽  
Constância Ayres ◽  
Lisa Reimer
Keyword(s):  
Aedes Aegypti ◽  
Insecticide Resistance ◽  
Vector Competence ◽  
Resistance Mechanisms ◽  
Target Site ◽  
Metabolic Resistance ◽  
Resistance Selection ◽  
Similar Phenotype ◽  
Target Site Resistance ◽  
Homozygous Resistant

Background: Laboratory reared mosquito colonies are essential tools to understand insecticide action. However, they differ considerably from wild populations and from each other depending on their origin and rearing conditions, which makes studying the effects of specific resistance mechanisms difficult. This paper describes our methods for establishing multiple resistant strains of Aedes aegypti from two colonies as a new resource for further research on metabolic and target site resistance. Methods: Two resistant colonies of Ae. aegypti, from Cayman and Recife, were selected through 10 generations of exposure to insecticides including permethrin, malathion and temephos, to yield eight strains with different profiles of resistance due to either target site or metabolic resistance. Resistance ratios for each insecticide were calculated for the selected and unselected strains. The frequency of kdr alleles (F1534C and V1016I) in the Cayman strains was determined using TaqMan assays. A comparative gene expression analysis among Recife strains was conducted using qPCR in larvae (CCae3A, CYP6N12, CYP6F3, CYP9M9) and adults (CCae3A, CYP6N12, CYP6BB2, CYP9J28a). Results: In the selected strain of Cayman, mortality against permethrin reduced almost to 0% and kdr became fixated by 5 generations. A similar phenotype was seen in the unselected homozygous resistant colony, whilst mortality in the susceptible homozygous colony rose to 82.9%. The Recife strains showed different responses between exposure to adulticide and larvicide, with detoxification genes in the temephos selected strain staying similar to the baseline, but a reduction in detoxification genes displayed in the other strains. Conclusions: These selected strains, with a range of insecticide resistance phenotypes and genotypes, will support further research on the effects of target-site and/or metabolic resistance mechanisms on various life-history traits, behaviours and vector competence of this important arbovirus vector.

scholarly journals Genome-wide gene expression profiling reveals that cuticle alterations and P450 detoxification are associated with pyrethroid resistance in Anopheles arabiensis populations from Ethiopia

2018 ◽  
Author(s):  
Eba Alemayehu Simma ◽  
Wannes Dermauw ◽  
Vasileia Balabanidou ◽  
Simon Snoeck ◽  
Astrid Bryon ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Arabiensis ◽  
Pyrethroid Resistance ◽  
Resistance Mutation ◽  
Resistance Mechanisms ◽  
Target Site ◽  
Malaria Vectors ◽  
Metabolic Resistance ◽  
Genome Wide ◽  
Target Site Resistance

AbstractBACKGROUNDVector control is the main intervention in malaria control and elimination strategies. However, the development of insecticide resistance is one of the major challenges for controlling malaria vectors. Anopheles arabiensis populations in Ethiopia showed resistance against both DDT and the pyrethroid deltamethrin. Although a L1014F target-site resistance mutation was present in the voltage gated sodium channel of investigated populations, the levels of resistance and biochemical studies indicated the presence of additional resistance mechanisms. In this study, we used genome-wide transcriptome profiling by RNAseq to assess differentially expressed genes between three deltamethrin and DDT resistant An. arabiensis field populations (Tolay, Asendabo, Chewaka) and two susceptible strains (Sekoru and Mozambique).RESULTSBoth RNAseq analysis and RT-qPCR showed that a glutathione-S-transferase, gstd3, and a cytochrome P450 monooxygenase, cyp6p4, were significantly overexpressed in the group of resistant populations compared to the susceptible strains, suggesting that the enzymes they encode play a key role in metabolic resistance against deltamethrin or DDT. Furthermore, a gene ontology enrichment analysis showed that expression changes of cuticle related genes were strongly associated with insecticide resistance, although this did not translate in increased thickness of the procuticle.CONCLUSIONOur transcriptome sequencing of deltamethrin/DDT resistant An. arabiensis populations from Ethiopia suggests non-target site resistance mechanisms and pave the way for further investigation of the role of cuticle composition in resistance.

scholarly journals Susceptibility to insecticides and resistance mechanisms in three populations of Aedes aegypti from Peru

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Jesus Pinto ◽  
Miriam Palomino ◽  
Leonardo Mendoza-Uribe ◽  
Carmen Sinti ◽  
Kelly A. Liebman ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Insecticide Resistance ◽  
Disease Transmission ◽  
Molecular Mechanisms ◽  
Mosquito Control ◽  
Resistance Mechanisms ◽  
Activity Levels ◽  
Metabolic Resistance ◽  
Glutathione S Transferases ◽  
Target Site Resistance

Abstract Background Epidemics of dengue, chikungunya and Zika are a growing threat to areas where Aedes aegypti are present. The efficacy of chemical control of Ae. aegypti is threatened by the increasing frequency of insecticide resistance. The objective of this study was to determine the susceptibility status as well as the biochemical and molecular mechanisms underlying insecticide resistance in three populations of Ae. aegypti in high risk areas of dengue, chikungunya, and Zika in Peru. Methods Bioassays were conducted on adult Ae. aegypti to evaluate their susceptibility to insecticides used currently or historically for mosquito control in Peru, including six pyrethroids, three organophosphates and one organochlorine, in populations of Ae. aegypti from the districts of Chosica (Department of Lima), Punchana (Department of Loreto) and Piura (Department of Piura). Resistance mechanisms were determined by biochemical assays to assess activity levels of key detoxification enzyme groups (nonspecific esterases, multi-function oxidases, glutathione S-transferases and insensitive acetylcholinesterase). Real-time PCR assays were used to detect two kdr mutations (V1016I and F1534C) on the voltage-gated sodium channel gene. Results Resistance to DDT was detected in all three populations, and resistance to pyrethroids was detected in all populations except the population from Chosica, which still exhibited susceptibility to deltamethrin. Resistance to organophosphates was also detected, with the exception of populations from Punchana and Piura, which still demonstrated susceptibility to malathion. In general, no increase or alteration of activity of any enzyme group was detected. Both 1016I and 1534C alleles were detected in Punchana and Piura, while only the 1534C allele was detected in Chosica. Conclusions The results suggest that resistance to multiple classes of insecticides exist in areas important to Ae. aegypti-borne disease transmission in Peru. The F1534C mutation was present in all 3 populations and the V1016I mutation was present in 2 populations. To our knowledge, this is the first report of the presence of 1016I and 1534C in Ae. aegypti in Peru. The absence of highly elevated enzymatic activity suggests that target site resistance is a key mechanism underlying insecticide resistance in these populations, although further research is needed to fully understand the role of metabolic resistance mechanisms in these populations.

scholarly journals Insecticide resistance in Anopheles arabiensis populations from Dakar and its suburbs: role of target site and metabolic resistance mechanisms

2018 ◽  
Vol 17 (1) ◽  
Author(s):  
A. Kane Dia ◽  
O. Kalsom Guèye ◽  
E. Amadou Niang ◽  
S. Mocote Diédhiou ◽  
M. Demba Sy ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Arabiensis ◽  
Resistance Mechanisms ◽  
Target Site ◽  
Metabolic Resistance

scholarly journals Insecticide Resistance Profiling of Anopheles coluzzii and Anopheles gambiae Populations in the Southern Senegal: Role of Target Sites and Metabolic Resistance Mechanisms

Genes ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1403
Author(s):  
Oumou. K. Gueye ◽  
Magellan Tchouakui ◽  
Abdoulaye K. Dia ◽  
Mouhamed B. Faye ◽  
Amblat A. Ahmed ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Resistance Mechanisms ◽  
Target Site ◽  
World Health ◽  
Anopheles Coluzzii ◽  
Partial Recovery ◽  
Metabolic Resistance ◽  
Genes Expression ◽  
Main Target

The emergence and spread of insecticide resistance among the main malaria vectors is threatening the effectiveness of vector control interventions in Senegal. The main drivers of this resistance in the Anopheles gambiae complex (e.g., An. gambiae and Anopheles coluzzii) remains poorly characterized in Senegal. Here we characterized the main target site and metabolic resistances mechanisms among the An. gambiae and An. coluzzii populations from their sympatric and allopatric or predominance area in Senegal. Larvae and pupae of An. gambiae s.l. were collected, reared to adulthood, and then used for insecticides susceptibility and synergist assays using the WHO (World Health Organisation) test kits for adult mosquitoes. The TaqMan method was used for the molecular characterization of the main target site insecticide resistance mechanisms (Vgsc-1014F, Vgsc-1014S, N1575Y and G119S). A RT-qPCR (Reverse Transcriptase-quantitative Polymerase Chaine Reaction) was performed to estimate the level of genes expression belonging to the CYP450 (Cytochrome P450) family. Plasmodium infection rate was investigated using TaqMan method. High levels of resistance to pyrethroids and DDT and full susceptibility to organophosphates and carbamates where observed in all three sites, excepted a probable resistance to bendiocarb in Kedougou. The L1014F, L1014S, and N1575Y mutations were found in both species. Pre-exposure to the PBO (Piperonyl butoxide) synergist induced a partial recovery of susceptibility to permethrin and full recovery to deltamethrin. Subsequent analysis of the level of genes expression, revealed that the CYP6Z1 and CYP6Z2 genes were over-expressed in wild-resistant mosquitoes compared to the reference susceptible strain (Kisumu), suggesting that both the metabolic resistance and target site mutation involving kdr mutations are likely implicated in this pyrethroid resistance. The presence of both target-site and metabolic resistance mechanisms in highly pyrethroid-resistant populations of An. gambiae s.l. from Senegal threatens the effectiveness and the sustainability of the pyrethroid-based tools and interventions currently deployed in the country. The Kdr-west mutation is widely widespread in An. coluzzii sympatric population. PBO or Duo nets and IRS (Indoor Residual Spraying) with organophosphates could be used as an alternative measure to sustain malaria control in the study area.

scholarly journals Insecticide resistance status of Aedes aegypti in Bangladesh

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.

Resistance Status to Deltamethrin, Permethrin, and Temephos Along With Preliminary Resistance Mechanism in Aedes aegypti (Diptera: Culicidae) From Punjab, Pakistan

2019 ◽  
Vol 56 (5) ◽  
pp. 1304-1311 ◽  
Author(s):  
Hafiz Azhar Ali Khan ◽  
Waseem Akram
Keyword(s):  
Aedes Aegypti ◽  
Insecticide Resistance ◽  
Resistance Mechanism ◽  
Resistance Mechanisms ◽  
Target Site Resistance ◽  
Control Failure ◽  
Moderate Resistance ◽  
High Level ◽  
Nonsignificant Effect

Abstract The use of insecticides such as deltamethrin, permethrin, and temephos has been a primary tool to manage Aedes aegypti (Linnaeus) in Punjab province, Pakistan; however, recent reports of control failure necessitate monitoring insecticide resistance. For this reason, we evaluated 12 field strains of Ae. aegypti from Punjab for resistance against deltamethrin, permethrin, and temephos along with underlying resistance mechanisms. For deltamethrin, high level of resistance was observed in Rawalpindi, Faisalabad, Sheikhupura, Lahore, Pattoki, and Kasur strains (RRLC50 > 10-fold); moderate level of resistance in Sargodha, Gujranwala, and Sialkot strains (RRLC50 = 5- to 10-fold), and low level of resistance in Okara, Multan, and Sahiwal strains (RRLC50 < 5-fold). In the case of permethrin, high level of resistance was found in all the field strains, except the Okara strain that exhibited moderate resistance. For temephos, five field strains, viz. Faisalabad, Rawalpindi, Kasur, Lahore, and Gujranwala, showed high level of resistance; five strains, viz. Sheikhupura, Sialkot, Pattoki, Sahiwal, and Okara, showed moderate resistance, and two strains from Multan and Sargodha showed low resistance to temephos. Synergism bioassays implementing piperonyl butoxide and S,S,S-tributylphosphorotrithioate exhibited a nonsignificant effect on synergizing toxicity of deltamethrin and permethrin in all field strains except the Lahore strain, suggesting the possible role of target-site resistance mechanism. However, both synergists had a significant effect on synergizing toxicity of temephos in all field strains, suggesting the possibility of metabolic-based mechanism of insecticide resistance. In conclusion, the study confirmed the presence of resistance to deltamethrin, permethrin, and temephos in the studied field strains of Ae. aegypti from Punjab, Pakistan.

scholarly journals Insecticide resistance status and mechanisms in Aedes aegypti populations from Senegal

2021 ◽  
Vol 15 (5) ◽  
pp. e0009393
Author(s):  
Ndeye Marie Sene ◽  
Konstantinos Mavridis ◽  
El Hadji Ndiaye ◽  
Cheikh Tidiane Diagne ◽  
Alioune Gaye ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Insecticide Resistance ◽  
Vector Control ◽  
Pyrethroid Resistance ◽  
Control Strategies ◽  
Resistance Mechanisms ◽  
Northern Region ◽  
Target Site ◽  
Limited Information ◽  
Resistance Mutations

Aedes aegypti is the main epidemic vector of arboviruses in Africa. In Senegal, control activities are mainly limited to mitigation of epidemics, with limited information available for Ae. aegypti populations. A better understanding of the current Ae. aegypti susceptibility status to various insecticides and relevant resistance mechanisms involved is needed for the implementation of effective vector control strategies. The present study focuses on the detection of insecticide resistance and reveals the related mechanisms in Ae. aegypti populations from Senegal. Bioassays were performed on Ae. aegypti adults from nine Senegalese localities (Matam, Louga, Barkedji, Ziguinchor, Mbour, Fatick, Dakar, Kédougou and Touba). Mosquitoes were exposed to four classes of insecticides using the standard WHO protocols. Resistance mechanisms were investigated by genotyping for pyrethroid target site resistance mutations (V1016G, V1016I, F1534C and S989P) and measuring gene expression levels of key detoxification genes (CYP6BB2, CYP9J26, CYP9J28, CYP9J32, CYP9M6, CCEae3a and GSTD4). All collected populations were resistant to DDT and carbamates except for the ones in Matam (Northern region). Resistance to permethrin was uniformly detected in mosquitoes from all areas. Except for Barkédji and Touba, all populations were characterized by a susceptibility to 0.75% Permethrin. Susceptibility to type II pyrethroids was detected only in the Southern regions (Kédougou and Ziguinchor). All mosquito populations were susceptible to 5% Malathion, but only Kédougou and Matam mosquitoes were susceptible to 0.8% Malathion. All populations were resistant to 0.05% Pirimiphos-methyl, whereas those from Louga, Mbour and Barkédji, also exhibited resistance to 1% Fenitrothion. None of the known target site pyrethroid resistance mutations was present in the mosquito samples included in the genotyping analysis (performed in > 1500 samples). In contrast, a remarkably high (20-70-fold) overexpression of major detoxification genes wasobserved, suggesting that insecticide resistance is mostly mediated through metabolic mechanisms. These data provide important evidence to support dengue vector control in Senegal.

scholarly journals Insecticide resistance status of Aedes aegypti in Bangladesh

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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