The Anopheles gambiae detoxification chip: A highly specific microarray to study metabolic-based insecticide resistance in malaria vectors

Abstract Background The rapid spread of insecticide resistance in malaria vectors and the rebound in malaria cases observed recently in some endemic areas underscore the urgent need to evaluate and deploy new effective control interventions. A randomized control trial (RCT) was conducted with the aim to investigate the benefit of deploying complementary strategies, including indoor residual spraying (IRS) with pirimiphos-methyl in addition to long-lasting insecticidal nets (LLINs) in Diébougou, southwest Burkina Faso. Methods We measured the susceptibility of the Anopheles gambiae (s.l.) population from Diébougou to conventional insecticides. We further monitored the efficacy and residual activity of pirimiphos-methyl on both cement and mud walls using a laboratory susceptible strain (Kisumu) and the local An. gambiae (s.l.) population. Results An. gambiae (s.l.) from Diébougou was resistant to DDT, pyrethroids (deltamethrin, permethrin and alphacypermethrin) and bendiocarb but showed susceptibility to organophosphates (pirimiphos-methyl and chlorpyrimiphos-methyl). A mixed-effect generalized linear model predicted that pirimiphos-methyl applied on cement or mud walls was effective for 210 days against the laboratory susceptible strain and 247 days against the local population. The residual efficacy of pirimiphos-methyl against the local population on walls made of mud was similar to that of cement (OR = 0.792, [0.55–1.12], Tukey’s test p-value = 0.19). Conclusions If data on malaria transmission and malaria cases (as measured trough the RCT) are consistent with data on residual activity of pirimiphos-methyl regardless of the type of wall, one round of IRS with pirimiphos-methyl would have the potential to control malaria in a context of multi-resistant An. gambiae (s.l.) for at least 7 months.

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Phenotypic Insecticide Resistance in Anopheles gambiae (Diptera: Culicidae): Specific Characterization of Underlying Resistance Mechanisms Still Matters

Journal of Medical Entomology ◽

10.1093/jme/tjaa195 ◽

2020 ◽

Author(s):

Adandé A Medjigbodo ◽

Luc S Djogbenou ◽

Aubin A Koumba ◽

Laurette Djossou ◽

Athanase Badolo ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Resistance Mechanisms ◽

Malaria Vectors ◽

Effective Control ◽

Anopheles Coluzzii ◽

Anopheles Gambiae S.S ◽

Diagnostic Dose ◽

Pirimiphos Methyl ◽

Old Adult

Abstract An effective control of malaria vectors requires an extensive knowledge of mechanisms underlying the resistance-phenotypes developed by these vectors against insecticides. We investigated Anopheles gambiae mosquitoes from Benin and Togo for their intensity of insecticide resistance and we discussed the involvement of genotyped mechanisms in the resistance-phenotypes observed. Three- to five-day-old adult mosquitoes emerged from field and laboratory An. gambiae larvae were assayed using WHO tube intensity tests against various doses of deltamethrin: 1× (0.05%); 2× (0.1%); 5× (0.25%); 7.5× (0.375%) and those of pirimiphos-methyl: 0.5× (0.125%); 1× (0.25%). Members of An. gambiae complex were screened in field populations using polymerase chain reaction (PCR) assays. The presence of kdrR(1014F/1014S) and ace-1R(119S) mutations was also investigated using TaqMan and PCR-RFLP techniques, respectively. Anopheles gambiae from field were very resistant to deltamethrin, whereas KisKdr and AcerKdrKis strains displayed 100% mortality rates at 2× the diagnostic dose. In contrast, the field mosquitoes displayed a low resistance-intensity against 1× the diagnostic dose of pirimiphos-methyl, whereas AcerKis and AcerKdrKis strains showed susceptibility at 0.5× the diagnostic dose. Anopheles gambiae s.s., Anopheles coluzzii, and Anopheles arabiensis were identified. Allelic frequencies of kdrR (1014F) and ace-1R (119S) mutations in the field populations varied from 0.65 to 1 and 0 to 0.84, respectively. The field An. gambiae displayed high-resistance levels against deltamethrin and pirimiphos-methyl when compared with those of the laboratory An. gambiae-resistant strains. These results exhibit the complexity of underlying insecticide resistance mechanisms in these field malaria vectors.

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Insecticide susceptibility status in individual species of the Anopheles gambiae complex (Diptera: Culicidae) in an area of The Gambia where pyrethroid impregnated bednets are used extensively for malaria control

Bulletin of Entomological Research ◽

10.1017/s0007485300034301 ◽

1995 ◽

Vol 85 (2) ◽

pp. 229-234 ◽

Cited By ~ 8

Author(s):

J. Hemingway ◽

S.W. Lindsay ◽

G.J. Small ◽

M. Jawara ◽

F.H. Collins

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Malaria Control ◽

The Gambia ◽

Malaria Vectors ◽

Cross Resistance ◽

Individual Species ◽

Sentinel Site ◽

Malaria Control Programme ◽

Biochemical Assays

AbstractPyrethroid-impregnated bednets are being used nationwide in The Gambia. The future success of this malaria control programme depends partly on the vectors remaining susceptible to those insecticides used for treating the nets. The present study was carried out on the south bank of the river Gambia, during the first large scale trial of nets in this country. Thus this area represents a sentinel site for detecting insecticide resistance in local vectors. This study gives an example of how a system of early detection for resistance problems can be set up in a relatively complex situation where multiple vectors and non-vectors are present. Samples of the Anopheles gambiae complex were caught indoors using light traps in twelve villages used in the bednet study. In all villages A. gambiae sensu stricto Giles was the predominant member of the complex as determined using the rDNA-PCR diagnostic assay. Limited bioassays with DDT and permethrin, and biochemical assays for a range of insecticide resistance mechanisms suggest that the A. gambiae complex remains completely susceptible to all major classes of commonly used insecticides including pyrethroids. Biochemical assays suggest that a low frequency of DDT resistance may occur in A. melas Theobald. This is based on elevated glutathione S-transferase levels coupled with increased levels of DDT metabolism and does not involve cross-resistance to pyrethroids. Therefore we do not envisage a decline in the efficacy of treated nets against malaria vectors in the study area in the immediate future, although monitoring should be continued whilst wide-scale use of impregnated bednets is operational.

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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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Identification of a rapidly-spreading triple mutant for high-level metabolic insecticide resistance in Anopheles gambiae provides a real-time molecular diagnostic for anti-malarial intervention deployment

10.1101/2021.02.11.429702 ◽

2021 ◽

Author(s):

Harun Njoroge ◽

Arjen van’t Hof ◽

Ambrose Oruni ◽

Dimitra Pipini ◽

Sanjay C. Nagi ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Pyrethroid Resistance ◽

Border Region ◽

Malaria Vectors ◽

Molecular Diagnostic ◽

Triple Mutant ◽

Pyrethroid Insecticides ◽

High Level ◽

Mutant Haplotype

AbstractInsecticide resistance provides both an increasingly pressing threat to the control of vector-borne diseases and insights into the remarkable capacity of natural populations to show rapid evolutionary responses to contemporary selection. Malaria control remains heavily dependent on deployment of pyrethroid insecticides, primarily in long lasting insecticidal nets (LLINs), but resistance in the major malaria vectors has increased over the last 15 years in concert with dramatic expansion of LLIN distributions. Identifying genetic mechanisms underlying high-level resistance in mosquitoes, which may almost entirely overcome pyrethroid efficacy, is crucial for the development and deployment of potentially resistance-breaking tools. Using the Anopheles gambiae 1000 genomes (Ag1000g) data we identified a very recent selective sweep in mosquitoes from Uganda which localized to a cluster of cytochrome P450 genes, including some commonly implicated in resistance. Further interrogation revealed a haplotype involving a trio of mutations, a nonsynonymous point mutation in Cyp6p4 (I236M), an upstream insertion of a partial Zanzibar-like transposable element (TE) and a duplication of the Cyp6aa1 gene. The mutations appear to have originated recently in An. gambiae from the Kenya-Uganda border region around Lake Victoria, with stepwise replacement of the double-mutant (Zanzibar-like TE and Cyp6p4-236M) with the triple-mutant haplotype (including Cyp6aa1 duplication), which has spread into the Democratic Republic of Congo and Tanzania. The triple-mutant haplotype is strongly associated with increased expression of genes able to metabolise pyrethroids and is strongly predictive of resistance to pyrethroids most notably deltamethrin, a commonly-used LLIN insecticide. Importantly, there was increased mortality in mosquitoes carrying the triple-mutation when exposed to nets co-treated with the synergist piperonyl butoxide (PBO). Frequencies of the triple-mutant haplotype remain spatially variable within countries, suggesting an effective marker system to guide deployment decisions for limited supplies of PBO-pyrethroid co-treated LLINs across African countries. Duplications of the Cyp6aa1 gene are common in An. gambiae across Africa and, given the enzymes metabolic activity, are likely to be a useful diagnostic for high levels of pyrethroid resistance.

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Characterization of resistance profile (intensity and mechanisms) of Anopheles gambiae in three communes of northern Benin, West Africa

Malaria Journal ◽

10.1186/s12936-021-03856-2 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Casimir Dossou Kpanou ◽

Hermann W. Sagbohan ◽

Fortuné Dagnon ◽

Germain G. Padonou ◽

Razaki Ossè ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Anopheles Gambiae ◽

High Intensity ◽

Mortality Rates ◽

Detoxification Enzymes ◽

Malaria Vectors ◽

Malaria Vector Control ◽

Biochemical Tests ◽

Diagnostic Dose

Abstract Background The selection and the spread of insecticide resistance in malaria vectors to the main classes of insecticides used in vector control tools are a major and ongoing challenge to malaria vector control programmes. This study aimed to determine the intensity of vector resistance to insecticides in three regions of Benin with different agro-ecological characteristics. Methods Larvae of Anopheles gambiae sensu lato (s.l.) were collected from September to November 2017 in different larval sites in three northern Benin communes: Parakou, Kandi and Malanville. Two to five-day-old, non-blood-fed, female mosquitoes were exposed to papers impregnated with deltamethrin, permethrin and bendiocarb at dosages of 1 × the diagnostic dose, 5 × and 10 × to determine the intensity of resistance in these vectors. Molecular frequencies of the kdr L1014F and ace-1R G119S insecticide resistance mutations and levels of detoxification enzymes were determined for mosquitoes sampled at each study site. Results Resistance to pyrethroids (permethrin and deltamethrin) was recorded in all three communes with mortality rates below 60% using the diagnostic dose (1x). The results obtained after exposure of An. gambiae to permethrin 10 × were 99% in Kandi, 98% in Malanville and 99% in Parakou. With deltamethrin 10x, mortality rates were 100% in Kandi, 96% in Malanville and 73% in Parakou. For the diagnostic dose of bendiocarb, suspected resistance was recorded in the communes of Malanville (97%) and Kandi (94%) while sensitivity was observed in Parakou (98%).Using the 10 × dose, mortality was 98% in Kandi, 100% in Malanville and 99% in Parakou. The frequencies of the kdr L1014F allele varied between 59 and 83% depending on the sites and species of the An. gambiae complex, while the frequency of the ace-1R G119S gene varied between 0 and 5%. Biochemical tests showed high levels of oxidase and esterase activity compared to the susceptible colony strain of An. gambiae sensu stricto (Kisumu strain). Conclusion Anopheles gambiae showed a generalized loss of susceptibility to permethrin and deltamethrin but also showed moderate to high intensity of resistance in different regions of Benin. This high intensity of resistance is a potential threat to the effectiveness of vector control.

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Mesocosm Experiments to Quantify Predation of Mosquito Larvae by Aquatic Predators to Determine Potential of Ecological Control of Malaria Vectors in Ethiopia

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18136904 ◽

2021 ◽

Vol 18 (13) ◽

pp. 6904

Author(s):

Beekam Kebede Olkeba ◽

Peter L. M. Goethals ◽

Pieter Boets ◽

Luc Duchateau ◽

Teshome Degefa ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Anopheles Arabiensis ◽

Anopheles Mosquito ◽

Malaria Vectors ◽

Physical Habitat ◽

Additional Tool ◽

Anopheles Mosquitoes ◽

Ecological Control ◽

Anopheles Gambiae S.L

Malaria parasites are transmitted to humans by infectious female Anopheles mosquitoes. Chemical-insecticide-based mosquito control has been successful in reducing the burden of malaria. However, the emergence of insecticide resistance in malaria vectors and concerns about the effect of the chemicals on the environment, human health, and non-target organisms present a need for new or alternative vector control intervention tools. Biocontrol methods using aquatic invertebrate predators have emerged as a potential alternative and additional tool to control mosquito populations. Ecological control specifically makes use of species insights for improving the physical habitat conditions of competitors and predators of vectors. A first step towards this is to gain knowledge on the predation potential of several typically present macroinvertebrates. Hence, this study aimed at (1) examining the influence of the predation of hemipterans on the number of emerging adult mosquitoes and (2) detecting Anopheles mosquito DNA in the gut of those predators. The prey and predators were collected from a range of water bodies located in the Gilgel Gibe watershed, southwest Ethiopia. A semi-field study was carried out using mesocosms which were constructed using plastic containers mimicking the natural aquatic habitat of immature Anopheles mosquitoes. Adult mosquitoes that emerged from the mesocosms were collected using a mechanical aspirator. At the end of the experiment, predators were withdrawn from the mesocosms and identified to genus level. Polymerase Chain Reaction (PCR) was employed to identify sibling species of Anopheles gambiae s.l. and to detect Anopheles mosquito DNA in the gut of the predators. Data were analysed using R software. Giant water bugs (belostomatids) were the most aggressive predators of Anopheles larvae, followed by backswimmers (notonectids) and water boatmen (corixids). All female Anopheles gambiae s.l. emerged from the mesocosms were identified as Anopheles arabiensis. Anopheles arabiensis DNA was detected in the gut content of hemipteran specimens analysed from the three families. The number of the adult mosquitoes emerging from the mesocosms was affected by the presence of predators. The findings of this study provide evidence of the potential use of aquatic macroinvertebrate predators as biocontrol agents against immature Anopheles mosquitoes and their potential to be considered as a component of integrated vector management for insecticide resistance and the combined restoration of aquatic ecosystems via smart ecological engineering.

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

10.21203/rs.3.rs-17402/v1 ◽

2020 ◽

Author(s):

Majidah Hamid-Adiamoh ◽

Alfred Amambua-Ngwa ◽

Davis Nwakanma ◽

Umberto D’Alessandro ◽

Gordon A. Awandare ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Blood Meal ◽

Northern Ghana ◽

Malaria Vectors ◽

Glutathione S Transferase ◽

Metabolic Resistance ◽

Anopheles Gambiae S.L ◽

Outdoor Resting ◽

Resting Behavior

Abstract Background Selection pressure from continued exposure to insecticides drives the development of insecticide resistance and changes in resting behavior of malaria vectors, which may support residual transmission in several endemic settings. There is a need to understand how resistance drives changes in resting behavior within vector species. Here, we examined the association between insecticide resistance and resting behavior of Anopheles gambiae s.l. in Northern Ghana. Methods Adult mosquitoes were collected both indoors and outdoors from two communities using mouth aspirators and pit shelters. F1 progenies from a subset of mosquitoes were exposed to dichloro diphenyl trichloroethane (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 (oxidases) were measured from unexposed F1 progenies using microplate assays. Results Susceptibility of An. coluzzii to deltamethrin 24hr post-exposure was significantly higher in indoor (mortality=5%) than the outdoor (mortality=2.5%) populations (P=0.02). The 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). The 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 s.s significantly associated with outdoor-resting behavior (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 the outdoor mosquito population (3%).Conclusions These findings revealed higher phenotypic resistance in indoor than outdoor-resting mosquitoes, but genotypic and metabolic resistance levels were higher in outdoor than the indoor mosquito populations. However, 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. Indication that human and animal blood meal indices were more prevalent in indoor-resting mosquitoes was also shown. Continued monitoring of changes in resting behavior within An. gambiae s.l. populations is recommended.

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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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Behavioral Responses of Pyrethroid-resistant Anopheles Gambiae Mosquitoes to Insecticide-treated Bed Net

10.21203/rs.3.rs-314905/v1 ◽

2021 ◽

Author(s):

Maxwell G. Machani ◽

Eric Ochomo ◽

Fred Amimo ◽

Andrew K. Githeko ◽

Guiyun Yan ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Malaria Transmission ◽

Malaria Vectors ◽

Bed Net ◽

Anopheles Gambiae S.S ◽

Field Environment ◽

Host Seeking ◽

Seeking Behavior ◽

Vector Behavior

Abstract Background: Long-lasting insecticidal nets are an effective tool in reducing malaria transmission. However, with increasing insecticide resistance little is known about how physiologically resistant malaria vectors behave around a human-occupied bed net, despite their importance in malaria transmission. We assessed the host-seeking behavior of the major malaria vector Anopheles gambiae s.s, when an intact human-occupied treated bed net is in place, with respect to their insecticide resistance status under semi-field conditions. Methods: Pyrethroid resistant and susceptible colonies of female Anopheles gambiae s.s aged 3-5 days that have been bred in our insectary, were color-marked with fluorescent powder and released inside a semi-field environment housing a hut which was occupied by a human host. Inside the hut, the occupant slept under an insecticide-treated bed net trap or untreated bed net trap. The window exit trap was installed to catch mosquitoes exiting the hut. A prokopack aspirator was used to collect indoor and outdoor resting mosquitoes in the morning. Clay pots were placed outside the hut to collect mosquitoes resting outdoors. Results: The proportion of resistant mosquitoes caught in the treated bed net trap was higher 43% (95% CI= [40.6-45.3]) compared to the susceptible mosquitoes 28.3% (OR=1.445; P<0.00019). The proportion of susceptible mosquitoes caught in the untreated bed net trap was higher 51.3% (95% CI= [48.8-53.6]) compared to the treated bed net trap 28.3% (95% CI= [26.3-30.5]) (OR=2.65; P<0.0001). Resistant mosquitoes were less likely to exit the house when a treated bed net was present (5.2%; 95% CI= [4.2-6.4]) compared to the susceptible mosquitoes (11.5%; 95% CI= [9.6-12.6]). The proportion of susceptible mosquitoes avoiding contact with the treated bed net and caught resting indoors in the hut (53.8%) and outdoors (64.5%) was higher compared to the resistant mosquitoes (indoors: 46.2%, outdoor: 35.4%). The susceptible females were 2.3 times more likely to stay outdoors away from the treated bed net (OR=2.25; 95% CI= [1.7-2.9]; P<0.0001).Conclusion: The results show that in the presence of a treated net, the host-seeking performance was not altered for the resistant mosquitoes, unlike the susceptible females that were observed to exit the house and remained outdoors when a treated net was used. However, further investigations of the behavior of resistant mosquitoes under natural conditions should be undertaken to confirm these observations and improve the current intervention which are threatened by insecticide resistance and altered vector behavior.

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