Insecticide Resistance In Mosquitoes Research Articles

Efficacy of Pirikool® 300 CS used for indoor residual spraying on three different substrates in semi-field experimental conditions

BackgroundVector control using insecticides is a key prevention strategy against malaria. Unfortunately, insecticide resistance in mosquitoes threatens all progress in malaria control. In the perspective of managing this resistance, new insecticide formulations are being tested to improve the effectiveness of vector control tools.MethodsThe efficacy and residual activity of Pirikool® 300 CS was evaluated in comparison with Actellic® 300 CS in experimental huts at the Tiassalé experimental station on three substrates including cement, wood and mud. The mortality, blood-feeding inhibition, exiting behaviour and deterrency of free-flying wild mosquitoes was evaluated. Cone bioassay tests with susceptible and resistant mosquito strains were conducted in the huts to determine residual efficacy.ResultsA total of 20,505 mosquitoes of which 10,979 (53%) wild female Anopheles gambiae were collected for 112 nights. Residual efficacy obtained from monthly cone bioassay was higher than 80% with the susceptible, laboratory-maintained An. gambiae Kisumu strain, from the first to the tenth study period on all three types of treated substrate for both Actellic® 300CS and Pirikool® 300CS. This residual efficacy on the wild Tiassalé strain was over 80% until the 4th month of study on Pirikool® 300CS S treated substrates. Overall 24-h mortalities of wild free-flying An. gambiae sensu lato which entered in the experimental huts over the 8-months trial on Pirikool® 300CS treatment was 50.5%, 75.9% and 52.7%, respectively, on cement wall, wood wall and mud wall. The positive reference product Actellic® 300CS treatment induced mortalities of 42.0%, 51.8% and 41.8% on cement wall, wood wall and mud wall.ConclusionPirikool® 300CS has performed really well against resistant strains of An. gambiae using indoor residual spraying method in experimental huts. It could be an alternative product for indoor residual spraying in response to the vectors' resistance to insecticides.

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti.

Insecticide resistance in mosquitoes is spreading worldwide and represents a growing threat to vector control. Insecticide resistance is caused by different mechanisms including higher metabolic detoxication, target-site modification, reduced penetration and behavioral changes that are not easily detectable with simple diagnostic methods. Indeed, most molecular resistance diagnostic tools are costly and labor intensive and then difficult to use for routine monitoring of insecticide resistance. The present study aims to determine whether mosquito susceptibility status against the pyrethroid insecticides (mostly used for mosquito control) could be established by the protein signatures of legs and/or thoraxes submitted to MALDI-TOF Mass Spectrometry (MS). The quality of MS spectra for both body parts was controlled to avoid any bias due to unconformity protein profiling. The comparison of MS profiles from three inbreeds Ae. aegypti lines from French Guiana (IRF, IR03, IR13), with distinct deltamethrin resistance genotype / phenotype and the susceptible reference laboratory line BORA (French Polynesia), showed different protein signatures. On both body parts, the analysis of whole protein profiles revealed a singularity of BORA line compared to the three inbreeding lines from French Guiana origin, suggesting that the first criteria of differentiation is the geographical origin and/or the breeding history rather than the insecticide susceptibility profile. However, a deeper analysis of the protein profiles allowed to identify 10 and 11 discriminating peaks from leg and thorax spectra, respectively. Among them, a specific peak around 4870 Da was detected in legs and thoraxes of pyrethroid resistant lines compared to the susceptible counterparts hence suggesting that MS profiling may be promising to rapidly distinguish resistant and susceptible phenotypes. Further work is needed to confirm the nature of this peak as a deltamethrin resistant marker and to validate the routine use of MS profiling to track insecticide resistance in Ae. aegypti field populations.

Evaluating the carnivorous efficacy of Utricularia aurea (Lamiales: Lentibulariaceae) on the larval stages of Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti (Diptera: Culicidae).

The emergence of insecticide resistance in mosquitoes necessitates the exploration and validation of sustainable biological strategies for controlling mosquitoes in their natural habitats. We assessed the predatory effect of Utricularia aurea Lour (Lamiales: Lentibulariaceae), an aquatic carnivorous plant found in the Indian subcontinent, Japan, and Australia, on 4 instars of Anopheles stephensi Liston, Culex quinquefasciatus Say, and Aedes aegypti Linn (Diptera: Culicidae), in the laboratory and field settings. In the laboratory setting, predation of larvae by U. aurea was highest during the first hour when it predated 45%, 61%, and 58% of first instars of An. stephensi, Cx. quinquefasciatus, and, Ae. aegypti, respectively, and, within 12 h, U. aurea preyed upon ~95% of the first, second, and third instars of the 3 mosquito species, ~80% of the fourth instars of An. stephensi and Ae. aegypti, and ~60% of fourth instars of Cx. quinquefasciatus. The predatory effect of U. aurea varied with mosquito species and instar. Broadly, predation risk declined with the increase of the instar size. In the field setting, at the end of 16 days, U. aurea predated 76% and 71% of the immature An. stephensi and Ae. aegypti, respectively. Our findings suggest U. aurea can be utilized as a potential biocontrol agent for controlling mosquito larvae in natural habitats; however, the current claim warrants additional investigations in a variety of natural habitats.

INSECTICIDE TOXICITY TO HONEY BEES: LESSONS LEARNED FROM STUDIES BY THE UNIVERSITY OF FLORIDA URBAN ENTOMOLOGY LABORATORY

Laboratory and field studies comparing the relative tolerance of mosquitoes (Aedes aegypti and Aedes albopictus) and non-target species, including the honey bee (Apis mellifera), and other non-target insects have shown that the non-target species can tolerate fairly well when exposed to insecticides used in mosquito control operations. Tolerance levels of non-target species vary due to factors such as the development of insecticide resistance in mosquitoes which require higher insecticide doses to reduce populations to adequate levels. However, well-planned mosquito control programs, can lower risks to honey bees and other non-target species. This review discusses the toxicity of mosquito control insecticides to the honey bee and other non-target insects.

Measuring insecticide resistance in a vacuum: exploring next steps to link resistance data with mosquito control efficacy.

Insecticide resistance is a great challenge facing mosquito operational control agencies across the United States, where few active ingredients with unique modes of action are available for use, increasing resistance pressure and further hampering resistance management strategies. Emergence and expansion of insecticide resistance in mosquitoes can be detected by resistance monitoring programs; however, there are gaps in our knowledge regarding the link between resistance bioassay results and operational control outcomes. Here, we review both public health and agricultural studies on pesticide resistance bioassays and control outcomes. A discussion on the main gaps in our knowledge of insecticide resistance and a review of resistance management practices is also presented. We conclude with research questions that can advance our understanding of resistance monitoring and control.

Larvicidal properties of terpenoid-based nanoemulsions against the dengue vector Aedes aegypti L. and their potential toxicity against non-target organism.

The development of insecticide resistance in mosquitoes of public health importance has encouraged extensive research into innovative vector control methods. Terpenes are the largest among Plants Secondary Metabolites and have been increasingly studied for their potential as insecticidal control agents. Although promising, terpenes are insoluble in water, and they show low residual life which limits their application for vector control. In this study, we developed and evaluated the performances of terpenoid-based nanoemulsions (TNEs) containing myrcene and p-cymene against the dengue vector Aedes aegypti and investigated their potential toxicity against non-target organisms. Our results showed that myrcene and p-cymene showed moderate larvicidal activity against mosquito larvae compared to temephos an organophosphate widely used for mosquito control. However, we showed similar efficacy of TNEs against both susceptible and highly insecticide-resistant mosquitoes from French Guyana, hence suggesting an absence of cross-resistance with conventional insecticides. We also showed that TNEs remained effective for up to 45 days in laboratory conditions. The exposure of zebrafish to TNEs triggered behavioral changes in the fish at high doses but they did not alter the normal functioning of zebrafish organs, suggesting a good tolerability of non-target organisms to these molecules. Overall, this study provides new insights into the insecticidal properties and toxicity of terpenes and terpenoid-based formulations and confirms that TNE may offer interesting prospects for mosquito control as part of integrated vector management.

Heat shock proteins, thermotolerance, and insecticide resistance in mosquitoes.

Mosquitoes transmit pathogens that pose a threat to millions of people globally. Unfortunately, widespread insecticide resistance makes it difficult to control these public health pests. General mechanisms of resistance, such as target site mutations or increased metabolic activity, are well established. However, many questions regarding the dynamics of these adaptations in the context of developmental and environmental conditions require additional exploration. One aspect of resistance that deserves further study is the role of heat shock proteins (HSPs) in insecticide tolerance. Studies show that mosquitoes experiencing heat stress before insecticide exposure demonstrate decreased mortality. This is similar to the observed reciprocal reduction in mortality in mosquitoes exposed to insecticide prior to heat stress. The environmental shifts associated with climate change will result in mosquitoes occupying environments with higher ambient temperatures, which could enhance existing insecticide resistance phenotypes. This physiological relationship adds a new dimension to the problem of insecticide resistance and further complicates the challenges that vector control and public health personnel face. This article reviews studies illustrating the relationship between insecticide resistance and HSPs or hsp genes as well as the intersection of thermotolerance and insecticide resistance. Further study of HSPs and insecticide resistance could lead to a deeper understanding of how environmental factors modulate the physiology of these important disease vectors to prepare for changing climatic conditions and the development of novel strategies to prevent vector-borne disease transmission.

Larvicidal potential of silver nanoparticles synthesized from Ocimum gratissimum leaf extracts against anopheles’ mosquito

Despite recent improvements in the methods used to eradicate and minimize mosquito populations, they still pose severe threats to human health. This study aimed to investigate the Larvicidal potentials of silver nanoparticles of Ocimum gratissimum on wild Anopheles larvae from Lagos. The plant used in this study was collected from the southeastern part of Nigeria, Ibite-Olo in Ezeagu Local Government Area of Enugu State. Wild Anopheles mosquito larvae were collected from a field at Kosofe Local Government Area of Lagos State, Nigeria where resistant Anopheline populations have been reported in and around Lagos State were brought to the laboratory and kept under standard insectary conditions (25–29 ℃;78–82% relative humidity). Aqueous extract of Ocimum gratissimum was made and subsequently into the silver nanoparticles. A bioassay test was performed according to WHO guidelines, with six different concentrations to assess the larvicidal activity and values obtained were subjected to log prohibit regression analysis to obtain LC50 and values with 95% confidence limit. The results of the larvicidal activities of the silver nanoparticles of Ocimum gratissimum showed that the mortality rate was dose-dependent with the percentage mortality increasing at a direct proportional rate as with dose and time of exposure. Ocimum gratissimum gave LC50 at 0.5 ppm and LC95 at 2.5 ppm respectively. The biosynthesized silver nanoparticles augmented the larvicidal activity of the silver nanoparticles of Ocimum gratissimum on wild Anopheles larvae from Lagos. Therefore, this study showed that the silver nanoparticles of Ocimum gratissimum have the potency to be used against an insecticide-resistant mosquito vector and are a suitable candidate for the development of novel natural insecticides.

Insecticide resistant mosquitoes remain thermal stress resistant, without loss of thermal plasticity

A major component of mosquito's climate change response is their heat tolerance, and any ability to rapidly adjust to extreme environmental conditions through phenotypic plasticity. The excessive use of insecticides for the control of major mosquito species leads to resistant populations, however it is largely unclear if this concurrently impacts thermal stress resistance and their potential to adjust tolerance via phenotypic plasticity. Culex pipiens pipiens, Culex pipiens molestus and Aedes albopictus populations obtained from the same region were subjected for 12 generations to selection trials to larvicides Diflubenzuron (DFB) and Bacillus thuringiensis subsp. israelensis (Bti) to develop insecticide resistance. Adults emerging from the selected populations were acclimated at different temperatures and the upper and lower critical thermal limits (CTmax and CTmin) were estimated using dynamic thermal assays. In addition, the supercooling points (SCPs) of non-acclimated adults of resistant and control populations were determined. Our results revealed marked differences in thermal response among the three species, the different acclimation regimes and sexes. Aedes albopictus was more resistant in high than low temperatures compared to both Culex pipiens biotypes. Culex forms responded similarly to heat but differently to cold stress. In both forms, females responded better than males to all thermal stressors. Acclimation at higher and lower temperatures improves CTmax and CTmin values, respectively in both insecticide resistant and control populations of all three species. Overall, selection to insecticides did not affect the thermal performance of adults. Hence, insecticide-resistant mosquito populations perform similarly to untreated ones and are capable of readily adapting to new environmental changes rising concerns regarding their geographic range expansion and disease transmission globally.

Predators as biocontrol agents of mosquito larvae in small and large habitats in Chiang Mai, Thailand.

Controlling mosquito-borne disease is a major global challenge due to the rise of insecticide-resistant mosquitoes. In response, we conducted a study in Chiang Mai Province, Thailand, which is one of the largest and the most popular cities for tourists in Southeast Asia, to explore the potential of local species as biological control agents for mosquito larvae. Mosquito larvae and aquatic predators were sampled from large and small habitats, while relevant physico-chemical parameters were measured. The study identified 560 predators and 1,572 mosquitoes, with most mosquito species belonging to the genus Culex. Additionally, the study identified 16 predator taxa, including four fish taxa and 12 taxa of predatory aquatic insects belonging to four orders: Coleoptera, Hemiptera, Odonata, and Diptera. The study found that several locally occurring predator species, namely Poecillia, Laccophilus, Lutzia, Toxorhynchites splendens, Agrionoptera, and Pseudarion, shared habitats with mosquitoes, indicating their potential as effective biological control agents for mosquito control. Conductivity, dissolved oxygen, and pH were the important physico-chemical parameters that affect both predators and mosquito larvae. Consequently, promoting native predators and reducing mosquito larvae through habitat management would be a sustainable and ecologically friendly approach in large habitats where it is not possible to remove mosquito oviposition sites. In smaller habitats, releasing local aquatic predators and removing oviposition sites may be a suitable strategy.

Promising Larvicidal Effects of Nanoliposomes Containing Carvone and Mentha spicata and Tanacetum balsamita Essential Oils Against Anopheles stephensi.

The use of synthetic pesticides to control the spread of mosquito-borne diseases has causedenvironmental pollution and insecticide resistance in mosquitoes. Developments of new green insecticides havethus received more attention to overcome these problems. Nanoliposomes containing carvone and essential oils were first prepared. The nanoliposomephysicochemical characteristics (particle size, morphology, and successful loading) were then evaluated byDynamic Light Scattering (DLS), Transmission Electron Microscopy (TEM), and the Attenuated Total Reflection-Fourier Transform InfraRed (ATR-FTIR) analyses. Larvicidal effects of carvone, Mentha spicata, and Tanacetumbalsamita essential oils were investigated against the main malaria vector, Anopheles stephensi, in non-formulatedand nanoformulated states. The larvicidal effects of nanoformulated states were significantly more potent (7.2 folds, 3.5 folds, and 8folds) than non-formulated states. Nanoliposomes containing M. spicata and T. balsamita essential oils with particlesizes of 175 ± 8 and 184 ± 5 nm showed the best efficacies (LC50 values = 9.74 and 9.36 μg/mL). The prepared samples could be used as new green potent larvicides against An stephensi mosquito infurther field trials. It is also recommended to investigate their efficacies against other mosquitoes.

Evolution of knockdown resistance (kdr) mutations of Aedes aegypti and Aedes albopictus in Hainan Island and Leizhou Peninsula, China.

Aedes aegypti and Aedes albopictus are important vectors of human arboviruses, transmitting arboviral diseases such as yellow fever, dengue, chikungunya and Zika. These two mosquitoes coexist on Hainan Island and the Leizhou Peninsula in China. Over the past 40 years, the distribution of Ae. albopictus has gradually expanded in these areas, while the distribution of Ae. aegypti has declined dramatically mainly due to the ecological changes and some other factors such as heavy use of insecticide indoor based on endophagic bloodfeeding of the species. This study focused on the knockdown resistance (kdr) genes of both mosquitoes, investigated their mutations, and analyzed their haplotype and evolutionary diversity combined with population genetic features based on the ND4/ND5 genes to further elucidate the molecular mechanisms underlying the development of insecticide resistance in both mosquitoes. Three mutations, S989P, V1016G and F1534C, were found to be present in Ae. aegypti populations, and the three mutations occurred synergistically. Multiple mutation types (F1534C/S/L/W) of the F1534 locus are found in Ae. albopictus populations, with the three common mutations F1534C, F1534S and F1534L all having multiple independent origins. The F1534W (TTC/TGG) mutation is thought to have evolved from the F1534L (TTC/TTG) mutation. The F1534S (TTC/TCG) mutation has evolved from the F1534S (TTC/TCC) mutation. The most common form of mutation at the F1534 locus found in this study was S1534C, accounting for 20.97%, which may have evolved from the F1534C mutation. In addition, a new non-synonymous mutation M1524I and 28 synonymous mutations were identified in Ae. albopictus populations. Correlation analysis showed that the genetic diversity of Ae. aegypti and Ae. albopictus populations did not correlate with their kdr haplotype diversity (P>0.05), but strong gene flow between populations may have contributed to the evolution of the kdr gene. The study of kdr gene evolution in the two mosquito species may help to identify the evolutionary trend of insecticide resistance at an early stage and provide a theoretical basis for improving the efficiency of biological vector control and subsequent research into new insecticides.

Quantitative proteomics analysis of permethrin and temephos-resistant Ae. aegypti revealed diverse differentially expressed proteins associated with insecticide resistance from Penang Island, Malaysia.

Synthetic insecticides are the primary vector control method used globally. However, the widespread use of insecticides is a major cause of insecticide-resistance in mosquitoes. Hence, this study aimed at elucidating permethrin and temephos-resistant protein expression profiles in Ae. aegypti using quantitative proteomics. In this study, we evaluated the susceptibility of Ae. aegypti from Penang Island dengue hotspot and non-hotspot against 0.75% permethrin and 31.25 mg/l temephos using WHO bioassay method. Protein extracts from the mosquitoes were then analysed using LC-ESI-MS/MS for protein identification and quantification via label-free quantitative proteomics (LFQ). Next, Perseus 1.6.14.0 statistical software was used to perform differential protein expression analysis using ANOVA and Student's t-test. The t-test selected proteins with≥2.0-fold change (FC) and ≥2 unique peptides for gene expression validation via qPCR. Finally, STRING software was used for functional ontology enrichment and protein-protein interactions (PPI). The WHO bioassay showed resistance with 28% and 53% mortalities in adult mosquitoes exposed to permethrin from the hotspot and non-hotspot areas. Meanwhile, the susceptibility of Ae. aegypti larvae revealed high resistance to temephos in hotspot and non-hotspot regions with 80% and 91% mortalities. The LFQ analyses revealed 501 and 557 (q-value <0.05) differentially expressed proteins in adults and larvae Ae. aegypti. The t-test showed 114 upregulated and 74 downregulated proteins in adult resistant versus laboratory strains exposed to permethrin. Meanwhile, 13 upregulated and 105 downregulated proteins were observed in larvae resistant versus laboratory strains exposed to temephos. The t-test revealed the upregulation of sodium/potassium-dependent ATPase β2 in adult permethrin resistant strain, H15 domain-containing protein, 60S ribosomal protein, and PB protein in larvae temephos resistant strain. The downregulation of troponin I, enolase phosphatase E1, glucosidase 2β was observed in adult permethrin resistant strain and tubulin β chain in larvae temephos resistant strain. Furthermore, the gene expression by qPCR revealed similar gene expression patterns in the above eight differentially expressed proteins. The PPI of differentially expressed proteins showed a p-value at <1.0 x 10-16 in permethrin and temephos resistant Ae. aegypti. Significantly enriched pathways in differentially expressed proteins revealed metabolic pathways, oxidative phosphorylation, carbon metabolism, biosynthesis of amino acids, glycolysis, and citrate cycle. In conclusion, this study has shown differentially expressed proteins and highlighted upregulated and downregulated proteins associated with insecticide resistance in Ae. aegypti. The validated differentially expressed proteins merit further investigation as a potential protein marker to monitor and predict insecticide resistance in field Ae. aegypti. The LC-MS/MS data were submitted into the MASSIVE database with identifier no: MSV000089259.

Evaluation of "Caserotek" a low cost and effective artificial blood-feeding device for mosquitoes.

Entomological research studies on mosquito vector biology, vector competence, insecticide resistance, dispersal, and survival (using mark-release-recapture techniques) often rely on laboratory-reared mosquito colonies to produce large numbers of consistently reared, aged, and sized mosquitoes. We developed a low-cost blood feeding apparatus that supports temperatures consistent with warm blooded animals, using commonly available materials found in low resource environments. We compare our system ("Caserotek") to Hemotek and glass/membrane feeding methods. Two experiments were conducted with Aedes aegypti (Linnaeus 1762) and one with Anopheles darlingi (Root 1926) (Diptera: Culicidae); 3 replicates were conducted for each experiment. Aedes aegypti female mosquitoes were provided chicken blood once per week for 30 min (Experiment #1) for 14 days or 1 hour (Experiment #2) for 21 days. Anopheles darlingi were fed once for 1 hour (Experiment #3). Blood-feeding rates, survival rates, and egg production were calculated across replicates. Caserotek had a significantly higher 30-min engorgement rate (91.1%) than Hemotek (47.7%), and the glass feeder (29.3%) whereas for 1-hour feeding, Hemotek had a significantly lower engorgement rate than either of the other two devices (78% versus 91%). Thirty-day survival was similar among the feeding devices, ranging from 86% to 99%. Mean egg production was highest for the Caserotek feeder (32 eggs per female) compared to the glass feeder and Hemotek device (21-22 eggs per female). Our new artificial feeding system had significantly higher blood feeding rates than for more expensive artificial systems and was equivalent to other fitness parameters. Caserotek only requires the ability to boil water to maintain blood temperatures using a Styrofoam liner. It can be easily scaled up to large production facilities and used under austere conditions.

MOSQUITO RESISTANCE TO INSECTICIDES IN BULGARIA

Mosquitoes are a group of blood-sucking insects that plays a major role in disease transmission in both humans and animals. They can pose a heavy nuisance, or serve as vectors of numerous viruses and parasites on most continents, including Europe. The strategy for successful control of the mosquito populations relies on the use of insecticides of five different categories: organochlorines, organophosphates, carbamates, pyrethroids, and biopesticides. The extensive use of the first four classes has resulted in the development of resistance originating in various mechanisms. The main ones are metabolic resistance and genetic resistance, manifested by point mutations in the insecticide targets, resulting in limited binding of neurotoxic substances. Much data is available on mosquito insecticide resistance in Bulgaria between 1948 and 1990, but only regarding organochlorines, organophosphates and carbamates. There is no data on pyrethroid resistance, which should raise an alarm for both public health authorities and private pest control companies since the only insecticides registered for professional mosquito control nowadays are pyrethroids. Therefore, there is an urgent need for the use of modern multidisciplinary approaches to study the resistance of native and invasive species of mosquitoes to insecticides, which should be a key contribution to elucidating their role in the circulation of pathogens of humans and animals and optimizing methods for controlling their populations.

Relationship between insecticide resistance profiles in Anopheles gambiae sensu lato and agricultural practices in Côte d’Ivoire

BackgroundInsecticide-based malaria vector control is increasingly undermined due to the development of insecticide resistance in mosquitoes. Insecticide resistance may partially be related to the use of pesticides in agriculture, while the level and mechanisms of resistance might differ between agricultural practices. The current study aimed to assess whether phenotypic insecticide resistance and associated molecular resistance mechanisms in Anopheles gambiae sensu lato differ between agricultural practices.MethodsWe collected An. gambiae s.l. larvae in six sites with three different agricultural practices, including rice, vegetable and cocoa cultivation. We then exposed the emerging adult females to discriminating concentrations of bendiocarb (0.1%), deltamethrin (0.05%), DDT (4%) and malathion (5%) using the standard World Health Organization insecticide susceptibility test. To investigate underlying molecular mechanisms of resistance, we used multiplex TaqMan qPCR assays. We determined the frequency of target-site mutations, including Vgsc-L995F/S and Vgsc-N1570Y, and Ace1-G280S. In addition, we measured the expression levels of genes previously associated with insecticide resistance in An. gambiae s.l., including the cytochrome P450-dependent monooxygenases CYP4G16, CYP6M2, CYP6P1, CYP6P3, CYP6P4, CYP6Z1 and CYP9K1, and the glutathione S-transferase GSTe2.ResultsThe An. gambiae s.l. populations from all six agricultural sites were resistant to bendiocarb, deltamethrin and DDT, while the populations from the two vegetable cultivation sites were additionally resistant to malathion. Most tested mosquitoes carried at least one mutant Vgsc-L995F allele that is associated with pyrethroid and DDT resistance. In the cocoa cultivation sites, we observed the highest 995F frequencies (80–87%), including a majority of homozygous mutants and several in co-occurrence with the Vgsc-N1570Y mutation. We detected the Ace1 mutation most frequently in vegetable-growing sites (51–60%), at a moderate frequency in rice (20–22%) and rarely in cocoa-growing sites (3–4%). In contrast, CYP6M2, CYP6P3, CYP6P4, CYP6Z1 and CYP9K1, previously associated with metabolic insecticide resistance, showed the highest expression levels in the populations from rice-growing sites compared to the susceptible Kisumu reference strain.ConclusionIn our study, we observed intriguing associations between the type of agricultural practices and certain insecticide resistance profiles in the malaria vector An. gambiae s.l. which might arise from the use of pesticides deployed for protecting crops.Graphical

Can insecticide resistance increase malaria transmission? A genetics-epidemiology mathematical modeling approach.

The great successes recorded in the fight against malaria over the last two decades, resulting from the wide scale implementation of insecticide-based interventions in malaria-endemic areas, has prompted a renewed global effort to eradicate malaria. The widespread emergence of insecticide resistance in the population of adult female malaria mosquitoes is considered to pose a potential challenge to such effort. In this study, we address one of the key questions in malaria ecology, namely whether or not insecticide resistance increase malaria transmission. We developed a genetics-epidemiology modeling framework that incorporates a detailed genotype structure of the gene that confers insecticide resistance in mosquitoes, malaria epidemiology in mosquitoes and humans (stratified based on whether or not they are protected by Long-lasting insecticide-treated nets (LLINs) indoors), genotype-specific mosquito repellance property of LLINs and mosquito biting behavior (indoor and outdoor bites). Conditions for the existence and local asymptotic stability of the various disease-free equilibria (by genotype) of the resulting genetic-epidemiology model are derived. This study identifies four parameters of the model that play a crucial role on quantifying the impact of insecticide resistance on malaria transmission, namely the parameters related to the level of the dominance of the resistant allele in heterozygous mosquitoes, the coverage of long-lasting insecticidal nets in the community, the probability of endophilic mosquitoes to successfully take a bloodmeal indoors and the proportion of new adult mosquitoes that are endophilic. We showed that, depending on the values of these four identified parameters, insecticide resistance can increase, decrease, or have no effect on malaria transmission. Our simulations show that malaria eradication can indeed be achieved using the currently-available chemical insecticides, even in the wake of the prevailing widespread insecticide resistance in malaria-endemic areas, if the insecticide-based interventions implemented can result in the attainment of the optimal values of the four identified parameters in malaria-endemic areas.

Inference of selection pressures that drive insecticide resistance in Anopheles and Culex mosquitoes in Korea

Pyrethroids are primarily used for mosquito control in Korea. However, high frequencies of mutations conferring resistance to not only pyrethroids but also to other insecticides have been found in mosquito populations. This study aimed to examine the hypothesis that insecticides used outside of public health may play a role in selection. Briefly, the resistance mutation frequencies to three insecticide groups (pyrethroids, organophosphates, and cyclodienes) were estimated in two representative groups of mosquito species (Anopheles Hyrcanus Group and Culex pipiens complex). The relationship between these frequencies and the land-use status of the collection sites was investigated through multiple regression analysis. In the Anopheles Hyrcanus Group, the frequencies of both ace1 (organophosphate resistance) and rdl (cyclodiene resistance) mutations were positively correlated with ‘proximity to golf course’, possibly be due to the insecticides used for turf maintenance. They also showed positive correlations with field area and rice paddy area, respectively, suggesting the role of agricultural insecticides in the selection of these resistance traits. For the Cx. pipiens complex, the kdr (pyrethroid resistance), ace1, and rdl mutations were positively correlated with the residential area, field, and rice paddy, respectively. Therefore, pyrethroids used for public health could serve as a direct source of resistance selection pressure against kdr, whereas non-public health insecticides may pose primary selection pressure against the ace1 and rdl traits. The current findings suggest that the insecticides used in agriculture and the golf industry play a significant role in mosquito selection, despite variations in the extent of indirect selection pressure according to the mosquito groups and insecticide classes.

Pyrethroid-treated bed nets impair blood feeding performance in insecticide resistant mosquitoes

The blood feeding performance of female mosquitoes directly impacts their ability to transmit malaria. Yet their host seeking and blood feeding behaviours in the presence of insecticide-treated nets (ITNs) are still poorly understood. This work explores how both insecticide resistant and susceptible Anopheles gambiae s.l. mosquitoes interact with pyrethroid nets (PermaNet 2.0 or Olyset net) or an untreated net (UTN) while attempting to blood feed on a human arm. Regardless of mosquito resistance status, the ITNs did not efficiently prevent host searching but reduced blood feeding success by 34.1 (29.31–38.95) %. The Permanet and Olyset net reduced to 227.5 (208.19–246.77) sec and 235.9 (214.03–257.74) sec the average blood feeding duration from 369.9 (342.78–397.04) sec with the UTN. The ingested blood volume was on average 22% lower for all mosquitoes exposed to insecticide. When feeding through ITNs, the blood volume flow rate of the susceptible strain increased by 35%, but no significant difference was found in the resistant strain. Thus, whilst the presence of the insecticide in ITNs reduced mosquito blood feeding success and blood volume, the mosquito’s ability to respond by accelerating her rate of blood ingestion may further reduce the impact of ITNs on resistant mosquitoes.

Using fruit flies to delve into mosquito insecticide resistance

With more than 3,000 species and an almost ubiquitous presence, the economic importance of mosquitoes cannot be overemphasized. Anopheles mosquitoes are vectors for infectious diseases such as malaria – an endemic disease in tropical and sub-tropical regions of the world that infects more than 200 million people worldwide and causes over 400,000 deaths annually, with most casualties being infants or inhabitants of sub-Saharan Africa. The Aedes aegytpi and Culex quinquefasciatus species of mosquitoes are also vectors for arboviruses such as chikungunya virus, dengue virus, western equine encephalitis virus, and Zika virus. Consequently, insecticides are frequently used to stem the population of mosquitoes. Nevertheless, mosquito insecticide resistance has emerged as a major problem that has contributed to numerous failed eradication campaigns for the aforementioned diseases. In this mini-review, we expound on how fruit flies (Drosophila melanogaster) could be a complementary model system for studying mosquito insecticide resistance, with the ultimate goal of confirming any promising leads in mosquitoes.