Insecticide resistance status of Anopheles arabiensis in irrigated and non-irrigated areas in western Kenya

Abstract Background Malaria control in Kenya is based on case management and vector control using long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS). However, the development of insecticide resistance compromises the effectiveness of insecticide-based vector control programs. The use of pesticides for agricultural purposes has been implicated as one of the sources driving the selection of resistance. The current study was undertaken to assess the status and mechanism of insecticide resistance in malaria vectors in irrigated and non-irrigated areas with varying agrochemical use in western Kenya. Methods The study was carried out in 2018–2019 in Homa Bay County, western Kenya. The bioassay was performed on adults reared from larvae collected from irrigated and non-irrigated fields in order to assess the susceptibility of malaria vectors to different classes of insecticides following the standard WHO guidelines. Characterization of knockdown resistance (kdr) and acetylcholinesterase-inhibiting enzyme/angiotensin-converting enzyme (Ace-1) mutations within Anopheles gambiae s.l. species was performed using the polymerase chain reaction (PCR) method. To determine the agricultural and public health insecticide usage pattern, a questionnaire was administered to farmers, households, and veterinary officers in the study area. Results Anopheles arabiensis was the predominant species in the irrigated (100%, n = 154) area and the dominant species in the non-irrigated areas (97.5%, n = 162), the rest being An. gambiae sensu stricto. In 2018, Anopheles arabiensis in the irrigated region were susceptible to all insecticides tested, while in the non-irrigated region reduced mortality was observed (84%) against deltamethrin. In 2019, phenotypic mortality was decreased (97.8–84% to 83.3–78.2%). In contrast, high mortality from malathion (100%), DDT (98.98%), and piperonyl butoxide (PBO)-deltamethrin (100%) was observed. Molecular analysis of the vectors from the irrigated and non-irrigated areas revealed low levels of leucine-serine/phenylalanine substitution at position 1014 (L1014S/L1014F), with mutation frequencies of 1–16%, and low-frequency mutation in the Ace-1R gene (0.7%). In addition to very high coverage of LLINs impregnated with pyrethroids and IRS with organophosphate insecticides, pyrethroids were the predominant chemical class of pesticides used for crop and animal protection. Conclusion Anopheles arabiensis from irrigated areas showed increased phenotypic resistance, and the intensive use of pesticides for crop protection in this region may have contributed to the selection of resistance genes observed. The susceptibility of these malaria vectors to organophosphates and PBO synergists in pyrethroids offers a promising future for IRS and insecticide-treated net-based vector control interventions. These findings emphasize the need for integrated vector control strategies, with particular attention to agricultural practices to mitigate mosquito resistance to insecticides. Graphic abstract

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Insecticide Resistance Status of Anopheles Arabiensis in Irrigated and Non-irrigated Areas in Western Kenya

10.21203/rs.3.rs-139000/v1 ◽

2021 ◽

Author(s):

Pauline Orondo ◽

Steven Nyanjom ◽

Harrysone Atieli ◽

John Githure ◽

Benyl Ondeto ◽

...

Keyword(s):

Public Health ◽

Insecticide Resistance ◽

Vector Control ◽

Anopheles Arabiensis ◽

Indoor Residual Spraying ◽

Malaria Vectors ◽

Initial Development ◽

Western Kenya ◽

Phenotypic Resistance ◽

Irrigated Area

Abstract Background: Malaria control in Kenya is based on case management and vector control using long lasting insecticidal nets (LLINs) and indoor residual spraying (IRS). Irrigation practices maintain vector population and thus transmission during dry season. Development of insecticide resistance further compromises the effectiveness of insecticide-based vector control programs. The aim of this study was to assess the status and mechanism of insecticide resistance in malaria vectors in irrigated and non-irrigated areas in western Kenya and the contribution of public health interventions and agriculture to insecticide resistance. Methodology: The study was carried out in 2018–2019 in Homa Bay County, western Kenya. Anopheline larvae were collected in irrigated and non-irrigated fields, reared to F1 adults and 2-5 day-old female vector mosquitoes were subjected to standard WHO insecticide susceptibility tests. The test specimens were then screened for knock-down resistance, kdr alleles, and analyzed for presence of acetyl-cholinesterase inhibiting enzyme; angiotensin-converting enzyme (Ace-1) genes. All field-collected samples were preserved for species identification by polymerase chain reaction. To ascertain the probable cause of vector resistance to insecticides, a questionnaire was administered to farmers, households and veterinary officers in the study area to assess the use of public health and agricultural insecticides/pesticides.Results: Anopheles arabiensis was the only species tested in irrigated (100%, n=154) area and predominant species in the non-irrigated areas (97.5%, n= 162) and the rest were An. gambiae sensu stricto. In 2018, susceptibility was observed in the vector species in the irrigated area and phenotypic resistance in the non-irrigated area while in 2019, phenotypic resistance was observed from all areas However, susceptibility to malathion (mortality 100%), DDT (98.98%-100%) and PBO- deltamethrin (100%) was observed. Molecular analysis of the vectors from the irrigated and non-irrigated areas revealed low levels of leucine- serine/ phenylalanine substitution at position 1014 (L1014S/ L1014F) with a mutation frequencies of 1%-16%, and almost zero mutation in Ace-1R gene (0.7%). In addition to very high coverage of LLINs impregnated with pyrethroids and IRS with organophosphate insecticides, pyrethroids were the predominant chemical class in pesticides used for crop and animal protection.Conclusion: Extensive use of pyrethroids in agriculture and public health could have resulted in the initial development of insecticide resistance. The susceptibility of these malaria vectors to organophosphates and PBO synergist in pyrethroids offers a promising future for IRS and ITN based vector control interventions.

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Status of insecticide susceptibility in Anopheles arabiensis and detection of the knockdown resistance mutation (kdr) concerning agricultural practices from Northern Sudan state, Sudan

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00142-1 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

M. Y. Korti ◽

T. B. Ageep ◽

A. I. Adam ◽

K. B. Shitta ◽

A. A. Hassan ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Anopheles Arabiensis ◽

Resistance Mutation ◽

Indoor Residual Spraying ◽

Mortality Rates ◽

Malaria Vectors ◽

Bed Nets ◽

Knockdown Resistance ◽

Insecticide Susceptibility

Abstract Background Chemical control has been the most efficient method in mosquito control, the development of insecticide resistance in target populations has a significant impact on vector control. The use of agricultural pesticides may have a profound impact on the development of resistance in the field populations of malaria vectors. Our study focused on insecticide resistance and knockdown resistance (kdr) of Anopheles arabiensis populations from Northern Sudan, related to agricultural pesticide usage. Results Anopheles arabiensis from urban and rural localities (Merowe and Al-hamadab) were fully susceptible to bendiocarb 0.1% and permethrin 0.75% insecticides while resistant to DDT 4% and malathion 5%. The population of laboratory reference colony F189 from Dongola showed a mortality of 91% to DDT (4%) and fully susceptible to others. GLM analysis indicated that insecticides, sites, site type, and their interaction were determinant factors on mortality rates (P < 0.01). Except for malathion, mortality rates of all insecticides were not significant (P > 0.05) according to sites. Mortality rates of malathion and DDT were varied significantly (P < 0.0001 and P < 0.05 respectively) by site types, while mortality rates of bendiocarb and permethrin were not significant (P >0.05). The West African kdr mutation (L1014F) was found in urban and rural sites. Even though, the low-moderate frequency of kdr (L1014F) mutation was observed. The findings presented here for An. arabiensis showed no correlation between the resistant phenotype as ascertained by bioassay and the presence of the kdr mutation, with all individuals tested except the Merowe site which showed a moderate association with DDT (OR= 6 in allelic test), suggesting that kdr genotype would be a poor indicator of phenotypic resistance. Conclusion The results provide critical pieces of information regarding the insecticide susceptibility status of An. arabiensis in northern Sudan. The usage of the same pesticides in agricultural areas seemed to affect the Anopheles susceptibility when they are exposed to those insecticides in the field. The kdr mutation might have a less role than normally expected in pyrethroids resistance; however, other resistance genes should be in focus. These pieces of information will help to improve the surveillance system and The implication of different vector control programs employing any of these insecticides either in the treatment of bed nets or for indoor residual spraying would achieve satisfactory success rates.

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Resting behaviour of malaria vectors in a highland and a lowland site of western Kenya: Implication on malaria vector control measures

10.1101/815175 ◽

2019 ◽

Author(s):

Maxwell G. Machani ◽

Eric Ochomo ◽

Fred Amimo ◽

Jackline Kosgei ◽

Stephen Munga ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Malaria Vector ◽

Control Measures ◽

Malaria Vectors ◽

Western Kenya ◽

Sporozoite Rate ◽

Resting Behaviour ◽

Outdoor Resting ◽

Indoor And Outdoor

AbstractBackgroundUnderstanding the interactions between increased insecticide resistance in field malaria vector populations and the subsequent resting behaviour patterns is important for planning adequate vector control measures in a specific context and sustaining the current vector interventions. The aim of this study was to investigate the resting behavior, host preference and infection with Plasmodium falciparum sporozoites by malaria vectors in different ecological settings of western Kenya with different levels of insecticide resistance.MethodsIndoor and outdoor resting Anopheline mosquitoes were sampled during the dry and rainy seasons in Kisian (lowland site) and Bungoma (highland site), both in western Kenya. WHO tube bioassay was used to determine levels of phenotypic resistance of first generation offspring (F1 progeny) of malaria vectors resting indoors and outdoors to deltamethrin. PCR-based molecular diagnostics were used for mosquito speciation, genotype for resistance mutations and to determine specific host blood meal origins. Enzyme-linked Immunosorbent Assay (ELISA) was used to determine mosquito sporozoite infections.ResultsOverall, 3,566 female Anopheles mosquitoes were collected with Anopheles gambiae s.l [In Bungoma, An. gambiae s.s (90.9%), An arabiensis (7.6%) and in Kisian, An. gambiae s.s (38.9%), An. arabiensis (60.2%)] being the most abundant species (74.7%) followed by An. funestus s.l (25.3%). The majority of An. gambiae s.l (85.4 and 58%) and An. funestus (96.6 and 91.1%) were caught resting indoors in Bungoma and Kisian respectively.Vgsc-1014S was observed at a slightly higher frequency in An. gambiae s.s hereafter(An. gambiae) resting indoor than outdoor (89.7 vs 84.6% and 71.5 vs 61.1%) in Bungoma and Kisian respectively. For An. arabiensis, Vgsc-1014S was 18.2% indoor and outdoor (17.9%) in Kisian. In Bungoma, the Vgsc-1014S was only detected in An. arabiensis resting indoors with a frequency of 10%. The Vgsc-1014F mutation was only present in An. gambiae resting indoors from both sites, but at very low frequencies in Kisian compared to Bungoma (0.8 and 9.2% respectively. In Bungoma, the sporozoite rates for An. funestus, An. gambiae, and An. arabiensis resting indoors were 10.9, 7.6 and 3.4 % respectively. For outdoor resting, An. gambiae and An. arabiensis in Bungoma, the sporozoite rates were 4.7 and 2.9 % respectively.Overall, in Bungoma, the sporozoite rate for indoor resting mosquitoes was 8.6% and 4.2% for outdoors. In Kisian the sporozoite rate was 0.9% for indoor resting An. gambiae. None of the outdoor collected mosquitoes in Kisian tested positive for sporozoite infections.ConclusionThe study reports high densities of insecticide-resistant An. gambiae and An. funestus resting indoors and the persistence of malaria transmission indoors with high entomological inoculation rates (EIR) regardless of the use of Long-lasting insecticidal nets (LLINs). These findings underline the difficulties of controlling malaria vectors resting and biting indoors using the current interventions. Supplemental vector control tools and implementation of sustainable insecticide resistance management strategies are needed in western Kenya.

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Monitoring of Insecticide Resistance in Anopheles culicifacies in Twelve Districts of Madhya Pradesh, Central India (2017–2019)

Journal of Tropical Medicine ◽

10.1155/2022/4404027 ◽

2022 ◽

Vol 2022 ◽

pp. 1-8

Author(s):

Ashok K. Mishra ◽

Praveen K Bharti ◽

Gyan Chand ◽

Aparup Das ◽

Himanshu Jayswar ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Indoor Residual Spraying ◽

Central India ◽

Control Programme ◽

Madhya Pradesh ◽

Control Measures ◽

Malaria Vectors ◽

Malaria Vector Control ◽

The Impact

Background. Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) are malaria vector control measures used in India, but the development of insecticide resistance poses major impediments for effective vector control strategies. As per the guidelines of the National Vector Borne Disease Control Programme (NVBDCP), the study was conducted in 12 districts of Madhya Pradesh to generate data on insecticide resistance in malaria vectors. Methods. The susceptibility tests were conducted on adult An. culicifacies as per the WHO standard technique with wild-caught mosquitoes. The blood-fed female mosquitoes were exposed in 3 to 4 replicates on each occasion to the impregnated papers with specified discriminating dosages of the insecticides (DDT: 4%, malathion: 5%, deltamethrin: 0.05%, and alphacypermethrin: 0.05%), for one hour, and mortality was recorded after 24-hour holding. Results. An. culicifacies was found resistant to DDT 4% in all the 12 districts and malathion in 11 districts. The resistance to alphacypermethrin was also observed in two districts, and possible resistance was found to alphacypermethrin in seven districts and to deltamethrin in eight districts, while the vector was found susceptible to both deltamethrin and alphacypermethrin in only 3 districts. Conclusion. An. culicifacies is resistant to DDT and malathion and has emerging resistance to pyrethroids, alphacypermethrin, and deltamethrin. Therefore, regular monitoring of insecticide susceptibility in malaria vectors is needed for implementing effective vector management strategies. However, studies to verify the impact of IRS with good coverage on the transmission of disease are required before deciding on the change of insecticide in conjunction with epidemiological data.

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The Current Insecticide Resistance in Main Malaria Vector Anopheles arabiensis in Yemen

Journal of Tropical Medicine ◽

10.1155/2020/5625019 ◽

2020 ◽

Vol 2020 ◽

pp. 1-5

Author(s):

Zalalham Al-Koleeby ◽

Ahmed El Aboudi ◽

Mithaq Assada ◽

Mohamed Al-Hadi ◽

Mohammed Abdalr Ahman ◽

...

Keyword(s):

Insecticide Resistance ◽

Malaria Vector ◽

Anopheles Arabiensis ◽

Resistance Management ◽

Indoor Residual Spraying ◽

Malaria Vectors ◽

Test Procedures ◽

Phenotypic Resistance ◽

Lambda Cyhalothrin ◽

Susceptibility Tests

Control of malaria vectors in Yemen relies on both indoor residual spraying using carbamate (bendiocarb) and long-lasting pyrethroids-treated nets. This paper reports the results of studies conducted to monitor the insecticide resistance of the main malaria vector, Anopheles arabiensis, to the insecticides currently used in the vector control in four different locations. Susceptibility tests were performed following the WHO test procedures. Two pyrethroids (lambda-cyhalothrin 0.05% and deltamethrin 0.05%) and one carbamate (bendiocarb 0.1%) were tested at diagnostic doses (DD). The five-fold DD of lambda-cyhalothrin and deltamethrin (0.25%) were also used to yield information on the intensity of resistance. Besides, tests with synergists were performed to assess the involvement of detoxifying enzyme in the phenotypic resistance of the populations of An. arabiensis to pyrethroids. The results of the performed susceptibility bioassay showed that the vector is susceptible to bendiocarb and resistant to lambda-cyhalothrin and deltamethrin in the four studied areas. The pyrethroids resistance is solely metabolic. This information could help policy-makers to plan insecticide resistance management. Bendiocarb is still an effective insecticide in the form of IRS. Concerning LLINS, it would be interesting to assess their effectiveness, combining a pyrethroid with PBO for the control of the pyrethroid-resistant malaria vector.

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Comparative assessment of insecticide resistance phenotypes in two major malaria vectors, Anopheles funestus and Anopheles arabiensis in south-eastern Tanzania

Malaria Journal ◽

10.1186/s12936-020-03483-3 ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Polius G. Pinda ◽

Claudia Eichenberger ◽

Halfan S. Ngowo ◽

Dickson S. Msaky ◽

Said Abbasi ◽

...

Keyword(s):

Insecticide Resistance ◽

Malaria Transmission ◽

Anopheles Arabiensis ◽

Pyrethroid Resistance ◽

Anopheles Funestus ◽

Indoor Residual Spraying ◽

Sub Saharan Africa ◽

World Health ◽

Malaria Vectors ◽

South Eastern

Abstract Background Long-lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS) have greatly reduced malaria transmission in sub-Saharan Africa, but are threatened by insecticide resistance. In south-eastern Tanzania, pyrethroid-resistant Anopheles funestus are now implicated in > 80% of malaria infections, even in villages where the species occurs at lower densities than the other vector, Anopheles arabiensis. This study compared the insecticide resistance phenotypes between the two malaria vectors in an area where pyrethroid-LLINs are widely used. Methods The study used the World Health Organization (WHO) assays with 1×, 5× and 10× insecticide doses to assess levels of resistance, followed by synergist bioassays to understand possible mechanisms of the observed resistance phenotypes. The tests involved adult mosquitoes collected from three villages across two districts in south-eastern Tanzania and included four insecticide classes. Findings At baseline doses (1×), both species were resistant to the two candidate pyrethroids (permethrin and deltamethrin), but susceptible to the organophosphate (pirimiphos-methyl). Anopheles funestus, but not An. arabiensis was also resistant to the carbamate (bendiocarb). Both species were resistant to DDT in all villages except in one village where An. arabiensis was susceptible. Anopheles funestus showed strong resistance to pyrethroids, surviving the 5× and 10× doses, while An. arabiensis reverted to susceptibility at the 5× dose. Pre-exposure to the synergist, piperonyl butoxide (PBO), enhanced the potency of the pyrethroids against both species and resulted in full susceptibility of An. arabiensis (> 98% mortality). However, for An. funestus from two villages, permethrin-associated mortalities after pre-exposure to PBO only exceeded 90% but not 98%. Conclusions In south-eastern Tanzania, where An. funestus dominates malaria transmission, the species also has much stronger resistance to pyrethroids than its counterpart, An. arabiensis, and can survive more classes of insecticides. The pyrethroid resistance in both species appears to be mostly metabolic and may be partially addressed using synergists, e.g. PBO. These findings may explain the continued persistence and dominance of An. funestus despite widespread use of pyrethroid-treated LLINs, and inform new intervention choices for such settings. In short and medium-term, these may include PBO-based LLINs or improved IRS with compounds to which the vectors are still susceptible.

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Insecticide exposure impacts vector–parasite interactions in insecticide-resistant malaria vectors

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2014.0389 ◽

2014 ◽

Vol 281 (1786) ◽

pp. 20140389 ◽

Cited By ~ 31

Author(s):

Haoues Alout ◽

Innocent Djègbè ◽

Fabrice Chandre ◽

Luc Salako Djogbénou ◽

Roch Kounbobr Dabiré ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Competence ◽

Resistance Mechanisms ◽

Malaria Vectors ◽

Anopheles Gambiae S.S ◽

Strong Trend ◽

Insecticide Exposure ◽

The Impact ◽

Selection Of ◽

Selection Of Resistance

Currently, there is a strong trend towards increasing insecticide-based vector control coverage in malaria endemic countries. The ecological consequence of insecticide applications has been mainly studied regarding the selection of resistance mechanisms; however, little is known about their impact on vector competence in mosquitoes responsible for malaria transmission. As they have limited toxicity to mosquitoes owing to the selection of resistance mechanisms, insecticides may also interact with pathogens developing in mosquitoes. In this study, we explored the impact of insecticide exposure on Plasmodium falciparum development in insecticide-resistant colonies of Anopheles gambiae s.s. , homozygous for the ace-1 G119S mutation (Acerkis) or the kdr L1014F mutation (Kdrkis). Exposure to bendiocarb insecticide reduced the prevalence and intensity of P. falciparum oocysts developing in the infected midgut of the Acerkis strain, whereas exposure to dichlorodiphenyltrichloroethane reduced only the prevalence of P. falciparum infection in the Kdrkis strain. Thus, insecticide resistance leads to a selective pressure of insecticides on Plasmodium parasites, providing, to our knowledge, the first evidence of genotype by environment interactions on vector competence in a natural Anopheles–Plasmodium combination . Insecticide applications would affect the transmission of malaria in spite of resistance and would reduce to some degree the impact of insecticide resistance on malaria control interventions.

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Comparative assessment of insecticide resistance phenotypes in two major malaria vectors, Anopheles funestus and Anopheles arabiensis in south-eastern Tanzania

10.21203/rs.3.rs-19762/v3 ◽

2020 ◽

Author(s):

Polius Gerazi Pinda ◽

Claudia Eichenberger ◽

Halfan S Ngowo ◽

Dickson S Msaky ◽

Said Abbasi ◽

...

Keyword(s):

Insecticide Resistance ◽

Malaria Transmission ◽

Anopheles Arabiensis ◽

Pyrethroid Resistance ◽

Anopheles Funestus ◽

Indoor Residual Spraying ◽

Sub Saharan Africa ◽

World Health ◽

Malaria Vectors ◽

South Eastern

Abstract BackgroundLong-lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS) have greatly reduced malaria transmission in sub-Saharan Africa, but are threatened by insecticide resistance. In south-eastern Tanzania, pyrethroid-resistant Anopheles funestus are now implicated in > 80% of malaria infections, even in villages where the species occurs at lower densities than the other vector, Anopheles arabiensis. This study compared the insecticide resistance phenotypes between the two malaria vectors in an area where pyrethroid-LLINs are widely used.MethodsThe study used the World Health Organization (WHO) assays with 1×, 5× and 10× insecticide doses to assess levels of resistance, followed by synergist bioassays to understand possible mechanisms of the observed resistance phenotypes. The tests involved adult mosquitoes collected from three villages across two districts in south-eastern Tanzania and included four insecticide classes.FindingsAt baseline doses (1×), both species were resistant to the two candidate pyrethroids (permethrin and deltamethrin), but susceptible to the organophosphate (pirimiphos-methyl). Anopheles funestus, but not An. arabiensis was also resistant to the carbamate (bendiocarb). Both species were resistant to DDT in all villages except in one village where An. arabiensis was susceptible. Anopheles funestus showed strong resistance to pyrethroids, surviving the 5× and 10× doses, while An. arabiensis reverted to susceptibility at the 5× dose. Pre-exposure to the synergist, piperonyl butoxide (PBO), enhanced the potency of the pyrethroids against both species and resulted in full susceptibility of An. arabiensis (>98% mortality). However, for An. funestus from two villages, permethrin-associated mortalities after pre-exposure to PBO only exceeded 90% but not 98%.ConclusionsIn south-eastern Tanzania, where An. funestus dominates malaria transmission, the species also has much stronger resistance to pyrethroids than its counterpart, An. arabiensis, and can survive more classes of insecticides. The pyrethroid resistance in both species appears to be mostly metabolic and may be partially addressed using synergists, e.g. PBO. These findings may explain the continued persistence and dominance of An. funestus despite widespread use of pyrethroid-treated LLINs, and inform new intervention choices for such settings. In short and medium-term, these may include PBO-based LLINs or improved IRS with compounds to which the vectors are still susceptible.

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Experimental evolution supports the potential of neonicotinoid-pyrethroid combination for managing insecticide resistance in malaria vectors

Scientific Reports ◽

10.1038/s41598-021-99061-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Marius Gonse Zoh ◽

Jean-Marc Bonneville ◽

Jordan Tutagata ◽

Frederic Laporte ◽

Behi K. Fodjo ◽

...

Keyword(s):

Public Health ◽

Insecticide Resistance ◽

Vector Control ◽

Target Site ◽

Site Mutation ◽

Protein Variant ◽

Selection Signature ◽

The Impact ◽

Selection Of ◽

Selection Of Resistance

AbstractThe introduction of neonicotinoids for managing insecticide resistance in mosquitoes is of high interest as they interact with a biochemical target not previously used in public health. In this concern, Bayer developed a combination of the neonicotinoid clothianidin and the pyrethroid deltamethrin (brand name Fludora Fusion) as a new vector control tool. Although this combination proved to be efficient against pyrethroid-resistant mosquitoes, its ability to prevent the selection of pyrethroid and neonicotinoid resistance alleles was not investigated. In this context, the objective of this work was to study the dynamics and the molecular mechanisms of resistance of An. gambiae to the separated or combined components of this combination. A field-derived An. gambiae line carrying resistance alleles to multiple insecticides at low frequencies was used as a starting for 33 successive generations of controlled selection. Resistance levels to each insecticide and target site mutation frequencies were monitored throughout the selection process. Cross resistance to other public health insecticides were also investigated. RNA-seq was used to compare gene transcription variations and polymorphisms across all lines. This study confirmed the potential of this insecticide combination to impair the selection of resistance as compared to its two separated components. Deltamethrin selection led to the rapid enrichment of the kdr L1014F target-site mutation. Clothianidin selection led to the over-transcription of multiple cytochrome P450s including some showing high homology with those conferring neonicotinoid resistance in other insects. A strong selection signature associated with clothianidin selection was also observed on a P450 gene cluster previously associated with resistance. Within this cluster, the gene CYP6M1 showed the highest selection signature together with a transcription profile supporting a role in clothianidin resistance. Modelling the impact of point mutations selected by clothianidin on CYP6M1 protein structure showed that selection retained a protein variant with a modified active site potentially enhancing clothianidin metabolism. In the context of the recent deployment of neonicotinoids for mosquito control and their frequent usage in agriculture, the present study highlights the benefit of combining them with other insecticides for preventing the selection of resistance and sustaining vector control activities.

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Neonicotinoid and pyrethroid combination: A tool to manage insecticide resistance in malaria vectors? Insights from experimental evolution

10.1101/2021.06.09.447494 ◽

2021 ◽

Author(s):

Marius Gonse Zoh ◽

Jean-Marc Bonneville ◽

Jordan Tutagana ◽

Frederic Laporte ◽

Behi Kouadio Fodjo ◽

...

Keyword(s):

Public Health ◽

Insecticide Resistance ◽

Vector Control ◽

Target Site ◽

Cross Resistance ◽

Site Mutation ◽

Selection Signature ◽

The Impact ◽

Selection Of ◽

Selection Of Resistance

Background: The introduction of neonicotinoids for managing insecticide resistance in mosquitoes is of high interest as they interact with a biochemical target not previously used in public health. In this concern, Bayer developed a combination of the neonicotinoid clothianidin and the pyrethroid deltamethrin (brand name Fludora Fusion) as a new vector control tool. Although this combination proved to be efficient against pyrethroid-resistant mosquitoes, its ability to prevent the selection of pyrethroid and neonicotinoid resistance alleles was not investigated. In this context, the objective of this work was to study the dynamics and the molecular mechanisms of resistance of An. gambiae to the separated or combined components of this combination. A field-derived An. gambiae line carrying resistance alleles to multiple insecticides at low frequencies was used as a starting for 33 successive generations of controlled selection. Resistance levels to each insecticide and target site mutation frequencies were monitored throughout the selection process. Cross resistance to other public health insecticides were also investigated. RNA-seq was used to compare gene transcription variations and polymorphisms across all lines. Results: This study confirmed the potential of this insecticide combination to impair the selection of resistance as compared to its two separated components. Deltamethrin selection led to the rapid enrichment of the kdr L1014F target-site mutation while clothianidin selection led to the over-transcription of multiple cytochrome P450s including some showing high homology with the ones conferring neonicotinoid resistance in other insects. A strong selection signature associated with clothianidin selection was observed on a cytochrome P450 gene cluster previously associated with resistance. Within this cluster, the gene CYP6M1 showed the highest selection signature together with a transcription profile supporting a role in clothianidin resistance. Modelling the impact of point mutations selected by clothianidin on CYP6M1 protein structure suggested that the selection of variants affecting its active site can enhance clothianidin metabolism. Conclusions: In the context of the recent deployment of neonicotinoids for mosquito control and their frequent usage in agriculture, the present study highlights the benefit of combining them with other insecticides for preventing the selection of resistance and sustaining vector control activities.

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