Assessment of diet choice by the yellow fever mosquito Aedes aegypti

To investigate feeding‐related decisions in Aedes aegypti (L.), adults are presented with simple diets of paired gustatory stimuli conveying information concerning energy content, nutrient richness, osmotic balance and food toxicity in a two‐diet matrix assay. Assessment of mosquito gut contents indicates that both sexes accept single sugar diets in a dose‐dependent manner. When presented with a choice between two different yet equimolar sugar solutions, more individuals of both sexes accept the disaccharides, sucrose and trehalose, than the monosacharrides, fructose and glucose. The combination of pyranose and furanose sugars in solution, either physically associated (as in sucrose) or present as monomers (as glucose and fructose), is accepted over solutions containing a single sugar moiety. Using the two‐diet matrix assay, mosquito diet‐choice is also tested between two equimolar sucrose ‘driver’ solutions in which one is presented with various concentrations of another potential feeding cue ‘test’ compound (i.e. each of the 20 naturally‐occurring amino acids, sodium chloride, quinine or caffeine). Diet‐choice between the ‘driver’ sucrose‐only solution and the solution of the ‘driver’ sucrose containing a ‘test’ amino acid is influenced by sex, amino acid concentration and sucrose concentration. There is also an example of synergism between the diet components, leucine and sucrose. Mosquitoes demonstrate a dose‐dependent acceptance of sucrose‐only diets over sodium chloride‐containing sucrose when presented together. Interestingly, the sucrose‐only diet is accepted by more mosquitoes than all concentrations of the saline‐containing sucrose diets except those approximately isotonic to mosquito haemolymph, at which concentration mosquitoes show no clear choice between the diets. More individuals of both sexes accept sucrose‐only diets than the diets of caffeine‐containing sucrose in a dose‐dependent manner. Only females, however, respond to quinine‐containing sucrose diets and modulate this behaviour in relation to the energetic reward: more females imbibed quinine‐containing sucrose at the higher sucrose concentration (1 m ). A systematic characterization of diet selection behaviour of A. aegypti is presented for 27 putative feeding cues potentially involved in nectar/honeydew feeding. This study will be used as a basis from which to investigate further the mosquito's assessment of food quality and ultimately host choice.     

Movement of Aedes aegypti following a sugar meal and its implication in the development of control strategies in Durán,Ecuador

We evaluated how the presence of sugar sources impacted the distribution of Aedes aegypti in different habitats in Durán , Ecuador. Land cover and normalized difference vegetation index maps were used to guide a random point sampling routine to select study grids (30 m × 30 m) in low vegetation (LV) and high vegetation (HV). Five individual plants, at one home in the LV and HV grid, were treated with a different colored, non‐attractive, 60% sucrose solution to determine mosquito feeding and movement. Sugar alone is not attractive to mosquitoes, so spraying vegetation with a dyed sugar solution can be used for visual determination of sugar feeding. Outdoor collections using BG sentinel traps and indoor collections using aspirators were conducted at the treatment home and with collection points at 20, 40, and 60 m surrounding the treatment home for three consecutive days. A total of 3,245 mosquitoes in two genera, Aedes and Culex, was collected. The proportion of stained Ae. aegypti females was 56.8% (510/898) and 0% for males. For Culex, 63.9% (248/388) females and 36.1% (140/388) males were collected stained. Aedes aegypti and Culex spp. were found up to 60 m stained in both LV and HV grids. Significantly more stained females Ae. aegypti were found inside homes compared to females and males of Culex spp. in both habitats. This study identifies that outdoor sugar feeding is a common behavior of Ae. aegypti and can be targeted as a control strategy in urban habitats in Latin America.     

Essential oils enhance the toxicity of permethrin against Aedes aegypti and Anopheles gambiae

Insecticide resistance and growing public concern over the safety and environmental impacts of some conventional insecticides have resulted in the need to discover alternative control tools. Naturally occurring botanically‐based compounds are of increased interest to aid in the management of mosquitoes. Susceptible strains of Aedes aegypti (Linnaeus) (Diptera: Culicidae) and Anopheles gambiae (Meigen) (Diptera: Culicidae) were treated with permethrin, a common type‐I synthetic pyrethroid, using a discriminate dose that resulted in less than 50% mortality. Piperonyl butoxide (PBO) and 35 essential oils were co‐delivered with permethrin at two doses (2 and 10 µg) to determine if they could enhance the 1‐h knockdown and the 24‐h mortality of permethrin. Several of the tested essential oils enhanced the efficacy of permethrin equally and more effectively than piperonyl butoxide PBO, which is the commercial standard to synergize chemical insecticide like pyrethroids. PBO had a strikingly negative effect on the 1‐h knockdown of permethrin against Ae. aegypti, which was not observed in An. gambiae. Botanical essential oils have the capability of increasing the efficacy of permethrin allowing for a natural alternative to classic chemical synergists, like PBO.     

Insecticidal sugar baits for adult biting midges

The mixing of an insecticide with sugar solution creates an oral toxin or insecticidal sugar bait (ISB) useful for reducing adult insect populations. The ability of ISBs to kill the biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae), a vector of bluetongue virus, epizootic hemorrhagic disease and vesicular stomatitis viruses, was tested. The commercial insecticide formulations (percentage active ingredient) tested included bifenthrin, cyfluthrin, deltamethrin, permethrin, dinotefuran, imidacloprid, thiamethoxam and spinosad. Mortality rates were determined for various concentrations of commercial formulations (0.01, 0.05, 0.1, 1, 2 and 3%) and observed at 1, 4, 10 and 24 h post‐exposure to the ISB. In the first set of assays, laboratory‐reared midges were fed sugar ad libitum and then exposed to insecticide‐treated sugar solutions to measure mortality. The second assay assessed competitive feeding: midges were provided with a control sugar solution (10% sucrose) in one vial, and a sugar and insecticide solution in another. Pyrethroid treatments resulted in the greatest mortality in the first hour at the lowest concentrations and spinosad consumption resulted in the least mortality. Biting midges were not deterred from feeding on the 1% ISB solutions despite the presence of an insecticide‐free alternative source of sugar.     

Frequency of pyrethroid resistance in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Thailand

Thirty‐two Aedes aegypti populations collected throughout Thailand and five populations of Aedes albopictus from southern Thailand were subjected to standard WHO contact bioassays to assess susceptibility to three commonly used synthetic pyrethroids: permethrin, deltamethrin, and lambda‐cyhalothrin. A wide degree of physiological response to permethrin was detected in Ae. aegypti, ranging from 56.5% survival (Lampang, northern Thailand) to only 4% (Kalasin in northeastern and Phuket in southern Thailand). All 32 populations of Ae. aegypti were found to have evidence of incipient resistance (62.5%) or levels of survival deemed resistant (37.5%) to permethrin. Four populations of Ae. albopictus were found with incipient resistance (97 – 80% mortality) and one with resistance (< 80%) to permethrin. The majority of Ae. aegypti populations (68.7%) was susceptible (> 98% mortality) to deltamethrin, with incipient resistance (observed 97–82% mortality) in other localities. In contrast, all populations of Ae. aegypti were completely susceptible (100% mortality) to the recommended operational dosage of lambda‐cyhalothrin. All five populations of Ae. albopictus were found completely susceptible to both deltamethrin and lambda‐cyhalothrin. Evidence of defined incipient or resistance to synthetic pyrethroids mandates appropriate response and countermeasures to mitigate further development and spread of resistance. In light of these findings, we conclude that routine and comprehensive susceptibility monitoring of dengue mosquito vectors to synthetic pyrethroids should be a required component of resistance management policies and disease control activities.     

An in vitro bioassay for the quantitative evaluation of mosquito repellents against Stegomyia aegypti (=Aedes aegypti) mosquitoes using a novel cocktail meal

To assess the efficacy of new insect repellents, an efficient and safe in vitro bioassay system using a multiple‐membrane blood‐feeding device and a cocktail meal was developed. The multiple‐membrane blood‐feeding device facilitates the identification of new insect repellents by the high‐throughput screening of candidate chemicals. A cocktail meal was developed as a replacement for blood for feeding females of Stegomyia aegypti (=Aedes aegypti) (L.) (Diptera: Culicidae). The cocktail meal consisted of a mixture of salt, albumin and dextrose, to which adenosine triphosphate was added to induce engorging. Feeding rates of St. aegypti on the cocktail meal and pig blood, respectively, did not differ significantly, but were significantly higher than the feeding rate on citrate phosphate dextrose‐adenine 1 (CPDA‐1) solutions, which had been used to replace bloodmeals in previous repellent assays. Dose‐dependent biting inhibition rates were analysed using probit analysis. The RD₅₀ (the dose producing 50% repellence of mosquito feeding) values of DEET, citronella, carvacrol, geraniol, eugenol and thymol were 1.62, 14.40, 22.51, 23.29, 23.83 and 68.05 µg/cm², respectively.     

Efficiency of fungus‐impregnated black cloths combined with Imidacloprid for the control of adult Aedes aegypti (Diptera: Culicidae)

Entomopathogenic fungi are potential candidates for use in integrated vector management. However, efficient delivery systems for these fungi need to be investigated. It is known that adult mosquitoes are attracted to dark surfaces, and therefore, black cotton cloths impregnated with Metarhizium anisopliae alone or in combination with the insecticide imidacloprid (IMI) were tested under laboratory conditions. Black cloths impregnated with fungus were also tested in large‐cage trials under natural extradomicile conditions. Blood‐fed Rockefeller and wild‐type strain Aedes aegypti had higher levels of survival when compared with sucrose‐fed counterparts following exposure to fungus‐impregnated cloths. However, when blood‐fed A. aegypti were exposed to a combination of M. anisopliae + IMI, the survival rates were statistically equal to those of sucrose‐fed females. Large‐cage trials showed significant decreases in A. aegypti survival following a minimum 12 h exposure of the mosquitoes to fungus‐impregnated cloths. Increased exposure times results in further reductions in survival. The synergism between M. anisopliae and IMI resulted in reduced survival rates independent of feeding regime under laboratory conditions. Fungus‐impregnated cloths tested under simulated field conditions, considered to be unfavourable for fungal infection, resulted in significant reductions in adult A. aegypti survival. We are currently testing the combined use of fungi and insecticides against blood‐fed insects under simulated field conditions.

Significance and Impact of the Study

The use of fungus‐impregnated cotton cloths is a promising point source application method for the control of adult Aedes aegypti, and this strategy could be incorporated into an integrated vector management programme aiming to reduce the incidence of dengue fever.     

Evaluation of boric acid as toxic sugar bait against resistant Aedes aegypti mosquitoes

Current methods of broad area application of contact insecticides used in mosquito control are becoming less effective, primarily due to resistance within mosquito populations. New methods that can deliver ingestible insecticides are being investigated as a means to mitigate resistance. This study evaluated insecticide delivery through toxic sugar baits (TSB) and resulting mortality of susceptible and resistant strains of Aedes aegypti. Two Ae. aegypti strains were evaluated using a 1% boric acid TSB: the susceptible Orlando 1952 (ORL) strain and the resistant Puerto Rican (PR) strain. The TSB resulted in high mortality for both ORL and PR strain of Ae. aegypti. Average mortality of female mosquitoes given TSB was 90.8% for PR and 99.3% for ORL. Our study suggests that targeting resistant mosquitoes with ingestible insecticides through TSBs could be a viable alternative to current mosquito control strategies and should be considered when developing an integrated vector management program.     

Gene expression profiles of the Southern house mosquito Culex quinquefasciatus during exposure to permethrin

Insecticide resistance is a major obstacle to the management of disease‐vectoring mosquitoes worldwide. The genetic changes and detoxification genes involved in insecticide resistance have been extensively studied in populations of insecticide‐resistant mosquitoes, however few studies have focused on the resistance genes upregulated upon insecticide exposure and the possible regulation pathways involved in insecticide resistance. To characterize the changes in gene expression during insecticide exposure, and to investigate the possible connection of known regulation pathways with insecticide resistance, we conducted RNA‐Seq analysis of a highly permethrin‐resistant strain of Culex quinquefasciatus following permethrin exposure. Gene expression profiles revealed a total of 224 upregulated and 146 downregulated genes when compared to a blank acetone carrier treated control, respectively, suggesting that there were multiple, but specific genes involved in permethrin resistance. Functional enrichment analysis showed that the upregulated genes contained multiple detoxification genes including a glutathione S‐transferase and multiple cytochrome P450 genes, as well as several immune‐related genes, while the downregulated genes consisted primarily of proteases and carbohydrate metabolism and transport. Further analysis showed that permethrin exposure resulted in a decrease in the expression of serum storage proteins and likely represented a delay in the development of the fourth instar possibly due to a decrease in feeding. This effect was more pronounced in an insecticide‐resistant strain than in an insecticide‐susceptible strain and may represent a behavioral mechanism of insecticide resistance in Culex mosquitoes.     

Effect of body size and sugar meals on oviposition of the yellow fever mosquito,Aedes aegypti (Diptera: Culicidae)

The effects of dietary sugar and body size on the oviposition of Ae. aegypti were studied under laboratory conditions. In female mosquitoes provided with sugar, the start of maximum fecundity was significantly delayed and the oviposition period was longer than in females provided with water. The peak of oviposition was also delayed in sugar‐fed females. Large females oviposited more eggs per day than small females at maximum fecundity and during eight days of observations. Large females also visited significantly more water‐containing cups in their cages per day than small females at maximum fecundity. During the eight days of observations, large females and sugar‐fed females visited more water‐containing cups in their cages than water‐fed small females. Both large females and sugar‐fed females oviposited their eggs at sites higher above the water line than water‐fed small females. These results suggested that large and sugar‐fed female Ae. aegypti mosquitoes had more energy reserves and oviposited their eggs at higher sites, which would lead to a time lag in hatching.     

Identification and Characterisation of Aedes aegypti Aldehyde Dehydrogenases Involved in Pyrethroid Metabolism

Background

Pyrethroid insecticides, especially permethrin and deltamethrin, have been used extensively worldwide for mosquito control. However, insecticide resistance can spread through a population very rapidly under strong selection pressure from insecticide use. The upregulation of aldehyde dehydrogenase (ALDH) has been reported upon pyrethroid treatment. In Aedes aegypti, the increase in ALDH activity against the hydrolytic product of pyrethroid has been observed in DDT/permethrin-resistant strains. The objective of this study was to identify the role of individual ALDHs involved in pyrethroid metabolism.

Methodology/Principal Findings

Three ALDHs were identified; two of these, ALDH9948 and ALDH14080, were upregulated in terms of both mRNA and protein levels in a DDT/pyrethroid-resistant strain of Ae. aegypti. Recombinant ALDH9948 and ALDH14080 exhibited oxidase activities to catalyse the oxidation of a permethrin intermediate, phenoxybenzyl aldehyde (PBald), to phenoxybenzoic acid (PBacid).

Conclusions/Significance

ALDHs have been identified in association with permethrin resistance in Ae. aegypti. Characterisation of recombinant ALDHs confirmed the role of this protein in pyrethroid metabolism. Understanding the biochemical and molecular mechanisms of pyrethroid resistance provides information for improving vector control strategies.     

Insecticide susceptibility status in Mexican populations of Stegomyia aegypti (= Aedes aegypti): a nationwide assessment

In Mexico, mosquito vector‐borne diseases are of public health concern as a result of their impact on human morbidity and mortality. The use of insecticides against adult mosquitoes is one of the most common ways of controlling mosquito population densities. However, the use of these compounds has resulted in the development of insecticide resistance. The aim of this study was to estimate susceptibility to six pyrethroids, two carbamates and two organophosphates in Mexican populations of Stegomyia aegypti (Linnaeus, 1762) (= Aedes aegypti) (Diptera: Culicidae) mosquitoes. Bottle insecticide susceptibility tests, with 1 h exposure, were performed on adult mosquitoes from 75 localities across 28 states. At 30 min of exposure, the proportion of fallen mosquitoes was recorded. After 60 min of exposure, mosquitoes were recovered in non‐treated containers and mortality was determined at 24 h after the set‐up of the experiment. In general, the carbamate insecticides represented the most effective group in terms of the proportion of mosquitoes fallen at 30 min (72–100%) and 24‐h mortality (97–100%). High and widespread resistance to pyrethroids Types I and II and, to a lesser extent, to organophosphates was observed. Insecticide susceptibility among and within states was highly variable.     

Susceptibility of three Colombian Aedes aegypti Linnaeus (Diptera: Culicidae) populations to a Lysinibacillus sphaericus formulation

The increased incidence of vector-borne diseases and insecticide resistance in mosquitos constitute public health concerns in the tropics. Biological control is an effective alternative in the management of Aedes aegypti Linnaeus populations. Lysinibacillus sphaericus a bacterium proved to be harmless for non-target organisms has shown promising entomopathogenic activity. In Colombia, the control of A. aegypti using L. sphaericus has not been contemplated as part of vector control programmes. To assess the susceptibility of three A. aegypti populations to a L. sphaericus formulation consisting of vegetative cells of 2362 and III(3)7 strains, simulated-field bioassays were conducted in the municipalities of Ricaurte in Cundinamarca and Tauramena in Casanare, as well as in the municipal inspection of San Joaquin, Cundinamarca. Sixty larvae were deposited in a test device, which contained nine litres of chlorine-free tap water and 100 ml of the bacterial formulation (10⁹ CFU/ml). Six replicates were set-up for treatment with bacteria and for the control. Larvae mortality was recorded at 24 and 48 hr. The three A. aegypti populations were susceptible to L. sphaericus formulation with a mortality rate higher than 90% at 48 hr. The formulation of L. sphaericus comprised of vegetative cells of 2362 and III(3)7 strains showed promising results that could guarantee the implementation of this formulation to control of A. aegypti populations.     

The presence of knockdown resistance mutations reduces male mating competitiveness in the major arbovirus vector,Aedes aegypti

BackgroundThe development of insecticide resistance in mosquitoes can have pleiotropic effects on key behaviours such as mating competition and host-location. Documenting these effects is crucial for understanding the dynamics and costs of insecticide resistance and may give researchers an evidence base for promoting vector control programs that aim to restore or conserve insecticide susceptibility.Methods and findingsWe evaluated changes in behaviour in a backcrossed strain of Aedes aegypti, homozygous for two knockdown resistance (kdr) mutations (V1016G and S989P) isolated in an otherwise fully susceptible genetic background. We compared biting activity, host location behaviours, wing beat frequency (WBF) and mating competition between the backcrossed strain, and the fully susceptible and resistant parental strains from which it was derived. The presence of the homozygous kdr mutations did not have significant effects on blood avidity, the time to locate a host, or WBF in females. There was, however, a significant reduction in mean WBF in males and a significant reduction in estimated male mating success (17.3%), associated with the isolated kdr genotype.ConclusionsOur results demonstrate a cost of insecticide resistance associated with an isolated kdr genotype and manifest as a reduction in male mating success. While there was no recorded difference in WBF between the females of our strains, the significant reduction in male WBF recorded in our backcrossed strain might contribute to mate-recognition and mating disruption. These consequences of resistance evolution, especially when combined with other pleiotropic fitness costs that have been previously described, may encourage reversion to susceptibility in the absence of insecticide selection pressures. This offers justification for the implementation of insecticide resistance management strategies based on the rotation or alternation of different insecticide classes in space and time.     

Insecticide resistance in dengue vectors from hotspots in Selangor,Malaysia

BackgroundIn Malaysia, dengue remains a top priority disease and usage of insecticides is the main method for dengue vector control. Limited baseline insecticide resistance data in dengue hotspots has prompted us to conduct this study. The present study reports the use of a map on the insecticide susceptibility status of Aedes aegypti and Aedes albopictus to provide a quick visualization and overview of the distribution of insecticide resistance.Method and resultsThe insecticide resistance status of Aedes populations collected from 24 dengue hotspot areas from the period of December 2018 until June 2019 was proactively monitored using the World Health Organization standard protocol for adult and larval susceptibility testing was conducted, together with elucidation of the mechanisms involved in observed resistance. For resistance monitoring, susceptibility to three adulticides (permethrin, deltamethrin, and malathion) was tested, as well as susceptibility to the larvicide, temephos. Data showed significant resistance to both deltamethrin and permethrin (pyrethroid insecticides), and to malathion (organophosphate insecticide) in all sampled Aedes aegypti populations, while variable resistance patterns were found in the sampled Aedes albopictus populations. Temephos resistance was observed when larvae were tested using the diagnostic dosage of 0.012mg/L but not at the operational dosage of 1mg/L for both species.ConclusionThe present study highlights evidence of a potential threat to the effectiveness of insecticides currently used in dengue vector control, and the urgent requirement for insecticide resistance management to be integrated into the National Dengue Control Program.     

Insecticide resistance,associated mechanisms and fitness aspects in two Brazilian Stegomyia aegypti (= Aedes aegypti) populations

In Brazil, insecticide resistance in Stegomyia aegypti (= Aedes aegypti) (Diptera: Culicidae) populations to pyrethroids and to the organophosphate (OP) temephos is disseminated. Currently, insect growth regulators (IGRs) and the OP malathion are employed against larvae and adults, respectively. Bioassays with mosquitoes from two northeast municipalities, Crato and Aracaju, revealed, in both populations, susceptibility to IGRs and malathion (RR₉₅ ≤ 2.0), confirming the effectiveness of these compounds. By contrast, temephos and deltamethrin (pyrethroid) resistance levels were high (RR₉₅ > 10), which is consistent with the use of intense chemical control. In Crato, RR₉₅ values were > 50 for both compounds. Knock‐down‐resistant (kdr) mutants in the voltage‐gated sodium channel, the pyrethroid target site, were found in 43 and 32%, respectively, of Aracaju and Crato mosquitoes. Biochemical assays revealed higher metabolic resistance activity (esterases, mixed function oxidases and glutathione‐S‐transferases) at Aracaju. With respect to fitness aspects, mating effectiveness was equivalently impaired in both populations, but Aracaju mosquitoes showed more damaging effects in terms of longer larval development, decreased bloodmeal acceptance, reduced engorgement and lower numbers of eggs laid per female. Compared with mosquitoes in Crato, Aracaju mosquitoes exhibited lower OP and pyrethroid RR₉₅, increased activity of detoxifying enzymes and greater effect on fitness. The potential relationship between insecticide resistance mechanisms and mosquito viability is discussed.     

A novel multiple membrane blood‐feeding system for investigating and maintaining Aedes aegypti and Aedes albopictus mosquitoes

A novel multiple membrane blood‐feeding system for mosquitoes has been developed for the study and routine maintenance of Aedes aegypti L. and Aedes albopictus Skuse that require a meal of vertebrate blood to produce eggs. This blood‐feeding system uses cattle collagen sausage‐casing membrane to facilitate feeding. The efficiency of this blood‐feeding system was compared to a live mice blood source. We observed that Ae. aegypti that fed on pig whole blood had 89.7% (w/o ATP) and 90.7% (w/ ATP) blood‐feeding rates, which were not significantly different from the mice‐fed ones (98.0%). Ae. albopictus fed on pig whole blood (w/ ATP) had a success rate of 84.4%, which was significantly different from the mice‐fed mosquitoes (51.1%). The feeding rates did not differ between sausage‐casing membrane and Parafilm‐M®. The survival rate, fecundity, pupation, and pupal emergence rates of Aedes females fed on pig whole blood were not significantly different from the mice‐fed ones. The artificial blood feeder can be applied to replace live animals as blood sources. Considering that this simple, inexpensive, convenient, and efficient feeding device can be built with common laboratory materials for research on Aedes mosquitoes.     

Differential mortalities of dengue-infected Aedes aegypti: preliminary results

Two ecotypes of Aedes aegypti, Ae. ae. aegypti and Ae. ae. formosus, were experimentally infected with a dengue 2 virus to test (1) the inheritance of susceptibility, and (2) the impact of infection on survival and reproduction. Ae. ae. aegypti, the main vector involved in dengue epidemics, displayed higher mortalities than Ae. ae. formosus, the ancestral form, which is a forest-dwelling, less anthropophilic species confined to Africa. Differential mortalities were observed between females with disseminated infection and females without disseminated infection. Ae. ae. formosus females with disseminated infection showed an increase in survival rate and reproduction success. These results are discussed in the light of changes in resource allocation that may occur in infected females.     

Imidacloprid impairs the post‐embryonic development of the midgut in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti)

The mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) is a vector for the dengue and yellow fever viruses. As blood digestion occurs in the midgut, this organ constitutes the route of entry of many pathogens. The effects of the insecticide imidacloprid on the survival of St. aegypti were investigated and the sub‐lethal effects of the insecticide on midgut development were determined. Third instar larvae were exposed to different concentrations of imidacloprid (0.15, 1.5, 3.0, 6.0 and 15.0 p.p.m.) and survival was monitored every 24 h for 10 days. Midguts from imidacloprid‐treated insects at different stages of development were dissected and processed for analyses by transmission electron microscopy, immunofluorescence microscopy and terminal deoxynucleotidyl transferase dUTP nick‐end labelling (TUNEL) assays. Imidacloprid concentrations of 3.0 and 15.0 p.p.m. were found to affect midgut development similarly. Digestive cells of the fourth instar larvae (L4) midgut exposed to imidacloprid had more multilamellar bodies, abundantly found in the cell apex, and more electron‐lucent vacuoles in the basal region compared with those from untreated insects. Moreover, imidacloprid interfered with the differentiation of regenerative cells, dramatically reducing the number of digestive and endocrine cells and leading to malformation of the midgut epithelium in adults. The data demonstrate that imidacloprid can reduce the survival of mosquitoes and thus indicate its potentially high efficacy in the control of St. aegypti populations.