Evaluation of the interaction between insecticide resistance-associated genes and malaria transmission in Anopheles gambiae sensu lato in central Côte d’Ivoire

Abstract Background There is evidence that the knockdown resistance gene (Kdr) L1014F and acetylcholinesterase-1 gene (Ace-1R) G119S mutations involved in pyrethroid and carbamate resistance in Anopheles gambiae influence malaria transmission in sub-Saharan Africa. This is likely due to changes in the behaviour, life history and vector competence and capacity of An. gambiae. In the present study, performed as part of a two-arm cluster randomized controlled trial evaluating the impact of household screening plus a novel insecticide delivery system (In2Care Eave Tubes), we investigated the distribution of insecticide target site mutations and their association with infection status in wild An. gambiae sensu lato (s.l.) populations. Methods Mosquitoes were captured in 40 villages around Bouaké by human landing catch from May 2017 to April 2019. Randomly selected samples of An. gambiae s.l. that were infected or not infected with Plasmodium sp. were identified to species and then genotyped for Kdr L1014F and Ace-1R G119S mutations using quantitative polymerase chain reaction assays. The frequencies of the two alleles were compared between Anopheles coluzzii and Anopheles gambiae and then between infected and uninfected groups for each species. Results The presence of An. gambiae (49%) and An. coluzzii (51%) was confirmed in Bouaké. Individuals of both species infected with Plasmodium parasites were found. Over the study period, the average frequency of the Kdr L1014F and Ace-1R G119S mutations did not vary significantly between study arms. However, the frequencies of the Kdr L1014F and Ace-1R G119S resistance alleles were significantly higher in An. gambiae than in An. coluzzii [odds ratio (95% confidence interval): 59.64 (30.81–131.63) for Kdr, and 2.79 (2.17–3.60) for Ace-1R]. For both species, there were no significant differences in Kdr L1014F or Ace-1R G119S genotypic and allelic frequency distributions between infected and uninfected specimens (P > 0.05). Conclusions Either alone or in combination, Kdr L1014F and Ace-1R G119S showed no significant association with Plasmodium infection in wild An. gambiae and An. coluzzii, demonstrating the similar competence of these species for Plasmodium transmission in Bouaké. Additional factors including behavioural and environmental ones that influence vector competence in natural populations, and those other than allele measurements (metabolic resistance factors) that contribute to resistance, should be considered when establishing the existence of a link between insecticide resistance and vector competence. Graphical Abstract

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Malaria Transmission, Vector Diversity, and Insecticide Resistance at a Peri-Urban Site in the Forest Zone of Ghana

Frontiers in Tropical Diseases ◽

10.3389/fitd.2021.739771 ◽

2021 ◽

Vol 2 ◽

Author(s):

Yaw Akuamoah-Boateng ◽

Ruth C. Brenyah ◽

Sandra A. Kwarteng ◽

Patrick Obuam ◽

Isaac Owusu-Frimpong ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Malaria Transmission ◽

Urban Setting ◽

World Health ◽

Malaria Vectors ◽

Urban Site ◽

Human Landing Catch ◽

Anopheles Coluzzii ◽

Anopheles Gambiae S.S

IntroductionRecent surge of Anopheles resistance to major classes of World Health Organization (WHO)-approved insecticides globally necessitates the need for information about local malaria vector populations. It is believed that insecticide efficacy loss may lead to operational failure of control interventions and an increase in malaria infection transmission. We investigated the susceptibility levels of malaria vectors to all classes of WHO-approved vector control insecticides and described the dynamics of malaria transmission in a peri-urban setting.MethodsFit 3–5-day-old adults that emerged from Anopheles larvae collected from several different sites in the study area were subjected to the WHO bioassay for detecting insecticide resistance. The knockdown resistance gene (kdr) mutations within the vector populations were detected using PCR. Entomological inoculation rates were determined using the human landing catch technique and Plasmodium falciparum circumsporozoite ELISA.ResultsThe malaria vectors from the study area were resistant to all classes of insecticides tested. Out of the 284 Anopheles complex specimen assayed for the resistance study, 265 (93.30%) were identified as Anopheles gambiae s.s. The kdr gene was detected in 90% of the Anopheles gambiae s.s. assayed. In an area where Anopheles coluzzii resistance to insecticides had never been reported, the kdr gene was detected in 78% of the Anopheles coluzzii sampled. The entomological inoculation rate (EIR) for the dry season was 1.44 ib/m/n, whereas the EIR for the rainy season was 2.69 ib/m/n.ConclusionsThis study provides information on the high parasite inoculation rate and insecticide resistance of malaria vectors in a peri-urban community, which is critical in the development of an insecticide resistance management program for the community.

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Interaction Between Insecticide Resistance-Associated Genes and Malaria Transmission in Anopheles Gambiae S. L. During a Cluster-Randomized Controlled Trial of A “lethal House Lure” in Central Côte D’ivoire

10.21203/rs.3.rs-637056/v1 ◽

2021 ◽

Author(s):

Rosine Z. Wolie ◽

Alphonsine A. Koffi ◽

Ludovic P. Ahoua Alou ◽

Eleanore D. Sternberg ◽

Oulo N’Nan-Alla ◽

...

Keyword(s):

Randomized Controlled Trial ◽

Insecticide Resistance ◽

Anopheles Gambiae ◽

Malaria Transmission ◽

Controlled Trial ◽

Cluster Randomized Controlled Trial ◽

Sub Saharan Africa ◽

Randomized Controlled ◽

Cluster Randomized ◽

The Impact

Abstract Background There is evidence that the Kdr L1014F and Ace-1R G119S mutations involved in pyrethroid and carbamate resistance in Anopheles gambiae influence malaria transmission in sub Saharan Africa. This is likely due to changes in behavior, life history, vectorial competence and capacity. In the present study, performed as part of a two-armed cluster randomized controlled trial (CRT) evaluating the impact of household screening plus a novel insecticide delivery system (In2Care EaveTubes), we investigated the distribution of insecticide target site mutations and their association with the infection status in wild An. gambiae s.l populations. Methods Mosquitoes were captured in 40 villages around Bouaké by human landing catches (HLC), from May 2017 to April 2019. Randomly selected sample of infected and uninfected An.gambiae s.l. with Plasmodium sp. were identified to species and then genotyped for Kdr L1014F and Ace-1R G119S mutations using quantitative polymerase chain reaction (qPCR) assays. The frequencies of the two alleles were compared between An. coluzzii and An. gambiae and then between infected and uninfected groups for each species. Results The presence of An. gambiae (49 %) and An. coluzzii (51%) was confirmed in Bouaké. Both species seemed to transmit equally Plasmodium parasites. Over the study period, the average frequency of the Kdr L1014F and Ace-1R G119S mutations did not vary significantly between study arms. However, the frequency of the Kdr L1014F and Ace-1R G119S resistance alleles were significantly higher in An. gambiae than in An. coluzzii (OR [95%CI]: 59.64 [30.81-131.63] for Kdr and OR [95%CI]: 2.79 [2.17–3.60], for Ace-1R). For both species, there were no significant differences in Kdr L1014F or Ace-1R G119S genotypic and allelic frequency distribution between infected and uninfected specimens (p > 0.05). Conclusions Either alone or in combination, Kdr L1014F and Ace-1R G119S showed no significant association with Plasmodium infection in wild An.gambiae and An. coluzzii, demonstrating similar competence for Plasmodium transmission in Bouaké. Additional factors influencing competence in natural population and those outside allele measurements contributing to resistance should be consider when establishing link between insecticide resistance and vector competence.

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Genetic variation at the Cyp6m2 putative insecticide resistance locus in Anopheles gambiae and Anopheles coluzzii

10.1101/2020.11.12.378943 ◽

2020 ◽

Author(s):

Martin G. Wagah ◽

Petra Korlević ◽

Christopher Clarkson ◽

Alistair Miles ◽

Mara K. N. Lawniczak ◽

...

Keyword(s):

Genetic Variation ◽

Insecticide Resistance ◽

Anopheles Gambiae ◽

Metabolic Activity ◽

High Frequency ◽

Cross Resistance ◽

Resistance Locus ◽

Anopheles Coluzzii ◽

Metabolic Resistance ◽

Enzymatic Function

AbstractBackgroundThe emergence of insecticide resistance is a major threat to malaria control programmes in Africa, with many different factors contributing to insecticide resistance in its vectors, Anopheles mosquitoes. CYP6M2 has previously been recognized as an important candidate in cytochrome P450-mediated detoxification in Anopheles mosquitoes. As it has been implicated in resistance against pyrethroids, organochlorines and carbamates, its broad metabolic activity makes it a potential agent in insecticide cross-resistance. Currently, allelic variation within the Cyp6m2 gene remains unknown.ResultsHere, we use Illumina whole-genome sequence data from Phase 2 of the Anopheles gambiae 1000 Genomes Project (Ag1000G) to examine genetic variation in the Cyp6m2 gene across 16 populations in 13 countries comprising Anopheles gambiae and Anopheles coluzzii mosquitoes. We find 15 missense biallelic substitutions at high frequency (defined as >5% frequency in one or more populations), that fall into five distinct haplotype groups that carry the main high frequency variants: A13T, D65A, E328Q, Y347F, I359V and A468S. We examine whether these alleles show evidence of selection either through potentially modified enzymatic function or by being linked to variants that change the transcriptional profile of the gene. Despite consistent reports of Cyp6m2 upregulation and metabolic activity in insecticide resistant Anophelines, we find no evidence of directional selection occurring on these variants or on the haplotype clusters in which they are found.ConclusionOur results imply that emerging resistance associated with Cyp6m2 is potentially driven by distant regulatory loci such as transcriptional factors rather than by its missense variants, or that other genes are playing a more significant role in conferring metabolic resistance.

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Characterizing the molecular and metabolic mechanisms of insecticide resistance in Anopheles gambiae s.l. in Faranah, Guinea

10.1101/610998 ◽

2019 ◽

Cited By ~ 1

Author(s):

Caleb Stica ◽

Claire L. Jeffries ◽

Seth R. Irish ◽

Yaya Barry ◽

Denka Camara ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Insecticide Resistance ◽

Anopheles Gambiae ◽

Scale Up ◽

Indoor Residual Spraying ◽

Control Program ◽

Sub Saharan Africa ◽

Health Concern ◽

Global Public Health ◽

Metabolic Resistance

AbstractBackgroundIn recent years, the scale-up of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) has greatly reduced malaria transmission. However, malaria remains a global public health concern with the majority of disease burden in sub-Saharan Africa. Insecticide resistance is a growing problem among Anopheles vector populations, with potential implications for the continued effectiveness of available control interventions. Improved understanding of current resistance levels and underlying mechanisms is essential to design appropriate management strategies and to mitigate future selection for resistance.MethodsAnopheles gambiae s.l. mosquitoes were collected from three villages in Faranah Prefecture, Guinea and their levels of susceptibility to seven insecticides were measured using CDC resistance intensity bioassays. Synergist assays with piperonyl butoxide (PBO) were also undertaken to assess the role of elevated mixed-function oxidases in resistance. RNA was extracted from 563 individuals and PCR was performed on cDNA to determine vector species, presence of target site mutations (L1014F kdr, N1575Y and G119S Ace-1), Plasmodium falciparum infection, and relative expression of three metabolic genes (CYP6M2, CYP6P3 and GSTD3).ResultsIn Faranah, resistance to permethrin and deltamethrin was observed, as well as possible resistance to bendiocarb. All assayed vector populations were fully susceptible to alpha-cypermethrin, pirimiphos-methyl, clothianidin and chlorfenapyr. Plasmodium falciparum infection was detected in 7.3% (37/508) mosquitoes tested. The L1014F kdr mutation was found in 100% of a sub-sample of 60 mosquitoes, supporting its fixation in the region. The N1575Y mutation was identified in 20% (113/561) of individuals, with ongoing selection evidenced by significant deviations from Hardy-Weinberg equilibrium. The G119S Ace-1 mutation was detected in 62.1% (18/29) of mosquitoes tested and was highly predictive of bendiocarb bioassay survival. The metabolic resistance genes, CYP6M2, CYP6P3 and GSTD3, were found to be overexpressed in wild resistant and susceptible An. gambiae s.s. populations, compared to a susceptible G3 colony. Furthermore, CYP6P3 was significantly overexpressed in bendiocarb survivors, implicating its potential role in carbamate resistance in Faranah.ConclusionsIdentification of intense resistance to permethrin and deltamethrin in Faranah, is of concern, as the Guinea National Malaria Control Program (NMCP) relies exclusively on the distribution of pyrethroid-treated LLINs for vector control. Study findings will be used to guide current and future control strategies in the region.

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

Genes ◽

10.3390/genes11121403 ◽

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.

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Insecticide resistance in indoor and outdoor-resting Anopheles gambiae in Northern Ghana

10.21203/rs.3.rs-17402/v3 ◽

2020 ◽

Author(s):

Majidah Hamid-Adiamoh ◽

Alfred Amambua-Ngwa ◽

Davis Nwakanma ◽

Umberto D’Alessandro ◽

Gordon A. Awandare ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Northern Ghana ◽

Malaria Vectors ◽

Anopheles Coluzzii ◽

Glutathione S Transferase ◽

Metabolic Resistance ◽

Phenotypic Resistance ◽

Resting Behaviour ◽

Outdoor Resting

Abstract Background Selection pressure from continued exposure to insecticides drives development of insecticide resistance and changes in resting behaviour of malaria vectors. There is need to understand how resistance drives changes in resting behaviour within vector species. The association between insecticide resistance and resting behaviour of Anopheles gambiae sensu lato (s.l.) in Northern Ghana was examined. Methods F1 progenies from adult mosquitoes collected indoors and outdoors were exposed to DDT, deltamethrin, malathion and bendiocarb using WHO insecticide susceptibility tests. Insecticide resistance markers including voltage-gated sodium channel (Vgsc)- 1014F, Vgsc-1014S, Vgsc-1575Y, glutathione-S-transferase epsilon 2 (GSTe2)-114T and acetylcholinesterase (Ace1)-119S, as well as blood meal sources were investigated using PCR methods. Activities of metabolic enzymes, acetylcholine esterase (AChE), non-specific β-esterases, glutathione-S-transferase (GST) and monooxygenases were measured from unexposed F1 progenies using microplate assays. Results Susceptibility of Anopheles coluzzii to deltamethrin 24hr post-exposure was significantly higher in indoor (mortality=5%) than outdoor (mortality=2.5%) populations (P=0.02). Mosquitoes were fully susceptible to malathion (mortality: indoor=98%, outdoor=100%). Susceptibility to DDT was significantly higher in outdoor (mortality=9%) than indoor (mortality=0%) mosquitoes (P=0.006). Mosquitoes were also found with suspected resistance to bendiocarb but mortality was not statistically different (mortality: indoor=90%, outdoor=95%. P=0.30). Frequencies of all resistance alleles were higher in F1 outdoor (0.11-0.85) than indoor (0.04-0.65) mosquito populations, while Vgsc-1014F in F0 An. gambiae sensu stricto (s.s) was significantly associated with outdoor-resting behaviour (P=0.01). Activities of non-specific β-esterase enzymes were significantly higher in outdoor than indoor mosquitoes (Mean enzyme activity: Outdoor=: 1.70/mg protein; Indoor=1.35/mg protein. P<0.0001). AChE activity was also more elevated in outdoor (0.62/mg protein) than indoor (0.57/mg protein) mosquitoes but this was not significant (P=0.08). Human blood index (HBI) was predominantly detected in indoor (18%) than outdoor mosquito populations (3%). Conclusions The overall results did not establish that there was a significant preference of resistant malaria vectors to solely rest indoors or outdoors, but varied depending on the resistant alleles present. Phenotypic resistance was higher in indoor than outdoor-resting mosquitoes, but genotypic and metabolic resistance levels were higher in outdoor than the indoor populations. Continued monitoring of changes in resting behaviour within An. gambiae s.l. populations is recommended.

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Evolution of the Pyrethroids Target-Site Resistance Mechanisms in Senegal: Early Stage of the Vgsc-1014F and Vgsc-1014S Allelic Frequencies Shift

Genes ◽

10.3390/genes12121948 ◽

2021 ◽

Vol 12 (12) ◽

pp. 1948

Author(s):

Moussa Diallo ◽

Majidah Hamid-Adiamoh ◽

Ousmane Sy ◽

Pape Cheikh Sarr ◽

Jarra Manneh ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Pyrethroid Resistance ◽

Early Stage ◽

Resistance Mechanisms ◽

Sub Saharan Africa ◽

Malaria Vectors ◽

Malaria Control Programme ◽

Anopheles Coluzzii ◽

Resistance Mutations

The evolution and spread of insecticide resistance mechanisms amongst malaria vectors across the sub-Saharan Africa threaten the effectiveness and sustainability of current insecticide-based vector control interventions. However, a successful insecticide resistance management plan relies strongly on evidence of historical and contemporary mechanisms circulating. This study aims to retrospectively determine the evolution and spread of pyrethroid resistance mechanisms among natural Anopheles gambiae s.l. populations in Senegal. Samples were randomly drawn from an existing mosquito sample, collected in 2013, 2017, and 2018 from 10 sentinel sites monitored by the Senegalese National Malaria Control Programme (NMCP). Molecular species of An. gambiae s.l. and the resistance mutations at the Voltage-gated Sodium Channel 1014 (Vgsc-1014) locus were characterised using PCR-based assays. The genetic diversity of the Vgsc gene was further analyzed by sequencing. The overall species composition revealed the predominance of Anopheles arabiensis (73.08%) followed by An. gambiae s.s. (14.48%), Anopheles coluzzii (10.94%) and Anopheles gambiae–coluzii hybrids (1.48%). Both Vgsc-1014F and Vgsc-1014S mutations were found in all studied populations with a spatial variation of allele frequencies from 3% to 90%; and 7% to 41%, respectively. The two mutations have been detected since 2013 across all the selected health districts, with Vgsc-L1014S frequency increasing over the years while Vgsc-1014F decreasing. At species level, the Vgsc-1014F and Vgsc-1014S alleles were more frequent amongst An. gambiae s.s. (70%) and An. arabiensis (20%). The Vgsc gene was found to be highly diversified with eight different haplotypes shared between Vgsc-1014F and Vgsc-1014S. The observed co-occurrence of Vgsc-1014F and Vgsc-1014S mutations suggest that pyrethroid resistance is becoming a widespread phenomenon amongst malaria vector populations, and the NMCP needs to address this issue to sustain the gain made in controlling malaria.

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The Impact of Insecticide Pre-Exposure on Longevity, Feeding Succession, and Egg Batch Size of Wild Anopheles gambiae s.l.

Journal of Tropical Medicine ◽

10.1155/2020/8017187 ◽

2020 ◽

Vol 2020 ◽

pp. 1-8

Author(s):

Grace Msangi ◽

Moses I. Olotu ◽

Aneth M. Mahande ◽

Anitha Philbert ◽

Eliningaya J. Kweka ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Vector Competence ◽

Survival Rates ◽

Indoor Residual Spraying ◽

Susceptible Strain ◽

Batch Size ◽

Management Options ◽

Anopheles Gambiae S.L ◽

The Impact

Background. Insecticide resistance among the vector population is the main threat to existing control tools available. The current vector control management options rely on applications of recommended public health insecticides, mainly pyrethroids through long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS). Regular monitoring of insecticide resistance does not provide information on important factors that affect parasite transmission. Such factors include vector longevity, vector competence, feeding success, and fecundity. This study investigated the impacts of insecticide resistance on longevity, feeding behaviour, and egg batch size of Anopheles gambiae s.l. Method. The larval sampling was conducted in rice fields using a standard dipper (350 ml) and reared to adults in field insectary. A WHO susceptibility test was conducted using standard treated permethrin (0.75%) and deltamethrin (0.05%) papers. The susceptible Kisumu strain was used for reference. Feeding succession and egg batch size were monitored for all survivors and control. Results. The results revealed that mortality rates declined by 52.5 and 59.5% for permethrin and deltamethrin, respectively. The mortality rate for the Kisumu susceptible strain was 100%. The survival rates of wild An. gambiae s.l. was between 24 and 27 days. However, the Kisumu susceptible strain blood meal feeding was significantly higher than resistant colony (t = 2.789, df = 21, P = 0.011 ). Additionally, the susceptible An. gambiae s.s. laid more eggs than the resistant An.gambiae s.l. colony (Χ2 = 1366, df = 1, P ≤ 0.05 ). Conclusion. It can, therefore, be concluded that the wild An. gambiae s.l. had increased longevity, blood feeding, and small egg batch size compared to Kisumu susceptible colonies.

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A pyrethroïd-treated bed net increases host attractiveness for Anopheles gambiae s.s. carrying the kdr allele in a dual-choice olfactometer

10.1101/077552 ◽

2016 ◽

Cited By ~ 1

Author(s):

Angélique Porciani ◽

Malal Diop ◽

Nicolas Moiroux ◽

Tatiana Kadoke-Lambi ◽

Anna Cohuet ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Sub Saharan Africa ◽

Malaria Vectors ◽

Malaria Vector Control ◽

Anopheles Gambiae S.S ◽

Host Seeking ◽

Seeking Behavior ◽

Sub Saharan ◽

The Impact

AbstractThe use of long lasting insecticide nets (LLINs) treated with pyrethroïd is known for its major contribution in malaria control. However, LLINs are suspected to induce behavioral changes in malaria vectors, which may in turn drastically affect their efficacy against Plasmodium sp. transmission. In sub Saharan Africa, where malaria imposes the heaviest burden, the main malaria vectors are widely resistant to pyrethroïds, the insecticide family used on LLINs, which also threatens LLIN efficiency. There is therefore a crucial need for deciphering how insecticide-impregnated materials might affect the host-seeking behavior of malaria vectors in regards to insecticide resistance. In this study, we explored the impact of permethrin-impregnated net on the host attractiveness for Anopheles gambiae mosquitoes, either susceptible to insecticides, or carrying the insecticide resistance conferring allele kdr. Groups of female mosquitoes were released in a dual-choice olfactometer and their movements towards an attractive odor source (a rabbit) protected by insecticide-treated (ITN) or untreated nets (UTN) were monitored. Kdr homozygous mosquitoes, resistant to insecticides, were more attracted by a host behind an ITN than an UTN, while the presence of insecticide on the net did not affect the choice of susceptible mosquitoes. These results suggest that permethrin-impregnated net is detectable by malaria vectors and that the kdr mutation impacts their response to a LLIN protected host. We discuss the implication of these results for malaria vector control.

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Genetic variation at the Cyp6m2 putative insecticide resistance locus in Anopheles gambiae and Anopheles coluzzii

Malaria Journal ◽

10.1186/s12936-021-03757-4 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Martin G. Wagah ◽

◽

Petra Korlević ◽

Christopher Clarkson ◽

Alistair Miles ◽

...

Keyword(s):

Genetic Variation ◽

Insecticide Resistance ◽

Anopheles Gambiae ◽

Metabolic Activity ◽

High Frequency ◽

Cross Resistance ◽

Resistance Locus ◽

Anopheles Coluzzii ◽

Metabolic Resistance ◽

Enzymatic Function

Abstract Background The emergence of insecticide resistance is a major threat to malaria control programmes in Africa, with many different factors contributing to insecticide resistance in its vectors, Anopheles mosquitoes. CYP6M2 has previously been recognized as an important candidate in cytochrome P450-mediated detoxification in Anopheles. As it has been implicated in resistance against pyrethroids, organochlorines and carbamates, its broad metabolic activity makes it a potential agent in insecticide cross-resistance. Currently, allelic variation within the Cyp6m2 gene remains unknown. Methods Here, Illumina whole-genome sequence data from Phase 2 of the Anopheles gambiae 1000 Genomes Project (Ag1000G) was used to examine genetic variation in the Cyp6m2 gene across 16 populations in 13 countries comprising Anopheles gambiae and Anopheles coluzzii mosquitoes. To identify whether these alleles show evidence of selection either through potentially modified enzymatic function or by being linked to variants that change the transcriptional profile of the gene, hierarchical clustering of haplotypes, linkage disequilibrium, median joining networks and extended haplotype homozygosity analyses were performed. Results Fifteen missense biallelic substitutions at high frequency (defined as > 5% frequency in one or more populations) are found, which fall into five distinct haplotype groups that carry the main high frequency variants: A13T, D65A, E328Q, Y347F, I359V and A468S. Despite consistent reports of Cyp6m2 upregulation and metabolic activity in insecticide resistant Anophelines, no evidence of directional selection is found occurring on these variants or on the haplotype clusters in which they are found. Conclusion These results imply that emerging resistance associated with Cyp6m2 is potentially driven by distant regulatory loci such as transcriptional factors rather than by its missense variants, or that other genes are playing a more significant role in conferring metabolic resistance.

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