Comparison of the insecticide susceptibilities of laboratory strains of Aedes aegypti and Aedes albopictus

A susceptible strain of Aedes albopictus derived from the Gainesville strain (Florida, USA) was established in our laboratory. The larvicidal efficacies of the neurotoxic insecticides temephos, permethrin and the pure cis and trans-permethrin isomers and the microbial insecticide Bacillus thuringiensis israelensis (Bti) against Ae. albopictus were estimated and compared to a susceptible strain of Aedes aegypti. The larvicidal effect of insect growth regulator pyriproxyfen was also evaluated in both mosquito strains. The median lethal concentration/median emergency inhibition values for Ae. aegypti and Ae. albopictus, respectively, were: temephos, 3.058 and 6.632 ppb, permethrin, 3.143 and 4.933 ppb, cis-permethrin, 4.457 and 10.068 ppb, trans-permethrin, 1.510 and 3.883 ppb, Bti, 0.655 and 0.880 ppb and pyriproxyfen, 0.00774 and 0.01642 ppb. Ae. albopictus was more tolerant than Ae. aegypti to all six larvicides evaluated. The order of susceptibility for Ae. aegypti was pyriproxyfen > Bti > trans-permethrin > temephos > permethrin > cis-permethrin and for Ae. albopictus was pyriproxyfen > Bti > trans-permethrin > permethrin > temephos > cis-permethrin. Because both species can be found together in common urban, suburban and rural breeding sites, the results of this work provide baseline data on the susceptibility of Ae. albopictus to insecticides commonly used for controlling Ae. aegypti in the field.     

Characterization of deltamethrin resistance in field populations of Aedes aegypti in Thailand.

Five field collections of adult Aedes aegypti mosquitoes from different areas in Bangkok and Pathum Thani provinces were subjected to susceptibility tests against deltamethrin. Low levels of resistance were detected among all populations tested (RR50 = 8-17.2) compared to the susceptible strain, Bora (French Polynesia). Among the five populations tested, the BKH (Bang Khen, Bangkok) and PSC (Phasicharoen, Bangkok) populations showed a higher level of deltamethrin resistance than the other three populations (RR50 of BKH = 17.2, and of PSC= 13.6) and cross-resistance to DDT was observed in these strains. Biochemical analysis showed a significant elevation of mixed function oxidases enzyme activity in all populations. There was an elevation of non-specific esterases in all populations except BKL, and there was no consistent association of glutathione S-transferases with deltamethrin and DDT resistance, although not all populations were bioassayed for DDT. The partial cDNA sequence of the para-type voltage-dependent sodium channel (IIS4-IIS6) was determined for BKH and PSC populations. Common amino acid substitution, leucine to phenylalanine in the IIS6 region, found for insects including Anopheles gambiae was not found in either the BKH or the PSC populations. However, two other amino acid substitutions (proline substituted with serine at position 64 in the PSC population and leucine with phenylalanine at position 69 in the BKH population) were found in the IIS5-IIS6 inter-segment region sequenced. The role these substitutions play in target site resistance is uncertain at this time.     

Glutathione S-transferase and insecticide resistance in laboratory strains and field populations of Musca domestica

Glutathione S-transferase (GST) activity of 10 house fly laboratory strains and 21 field populations are described with two substrates, 1-chloro-2,4-dinitrobenzene (CDNB) and 3,4-dichloronitrobenzene (DCNB), commonly associated with resistance. Three laboratory strains selected by tetrachlorvinphos, lindane (gamma-HCH) and dimethoate, respectively, had significantly elevated CDNB- and DCNB-GST activities. The multiresistant field population 791a had significantly elevated CDNB- and DCNB-GST activities. Many strains recorded several individuals with high CDNB- and/or DCNB-GST activity and to evaluate these differences, phenotypes were defined by cluster analysis. A phenotype, GST-R, indicating high CDNB- and DCNB-GST activities, was found in 23 of 31 laboratory strains or field populations. GST-R was likely to be involved in gamma-HCH resistance in strain 17e, tetrachlorvinphos resistance in strain 39m2b, and dimethoate resistance in strain 49r2b. The frequency of GST-R in selected laboratory strains and field population correlated with the frequency of house flies surviving the organophosphate azamethiphos either topically applied or by ingestion. There was no significant correlation between GST activity and the toxicity of the organophosphate dimethoate or the pyrethroids bioresmethrin and pyrethrin.     

Association of insecticide use and alteration on Aedes aegypti susceptibility status

Dengue and dengue hemorrhagic fever, vector-borne diseases transmitted by the mosquito Aedes aegypti, are presently important public health problems in Brazil. As the strategy for disease control is based on vector control through the use of insecticides, the development of resistance is a threat to programs efficacy. The objective of this study was to compare the Aedes aegypti susceptibility in nine vector populations from the state of S?o Paulo and seven from Northeast region of Brazil, since there was a difference on group of insecticide used between the areas. Bioassays with larvae and adult were performed according to the World Health Organization methods. The results showed higher resistance levels to organophosphates group in populations from the Northeast region where this group was used for both larvae and adult control than in S?o Paulo where organophosphates were used for larvae and pyretroids for adult control. Resistance to pyretroids in adults was widespread in S?o Paulo after ten years of use of cypermethrin while in vector populations from the Northeast region it was punctual. The difference in resistance profile between the areas is in accordance to the group of insecticide used.     

Characterisation of DDT and pyrethroid resistance in Trinidad and Tobago populations of Aedes aegypti

Insecticide resistance is an important factor in the effectiveness of Aedes aegypti control and the related spread of dengue. The objectives of this study were to investigate the status of the organochlorine dichlorodiphenyltrichloroethane (DDT) and pyrethroid (permethrin and deltamethrin) resistance in Trinidad and Tobago populations of Ae. aegypti and the underlying biochemical mechanisms. Nine populations of Ae. aegypti larvae from Trinidad and Tobago were assayed to DDT and PYs using the Centers for Disease Control and Prevention (CDC) time-mortality-based bioassay method. A diagnostic dosage (DD) was established for each insecticide using the CAREC reference susceptible Ae. aegypti strain and a resistance threshold (RT), time in which 98-100% mortality was observed in the CAREC strain, was calculated for each insecticide. Mosquitoes which survived the DD and RT were considered as resistant, and the resistance status of each population was categorised based on the WHO criteria with mortality <80% indicative of resistance. Biochemical assays were conducted to determine the activities of α and β esterases, mixed function oxidases (MFO) and glutathione-S-transferases (GST) enzymes which are involved in resistance of mosquitoes to DDT and PYs. Enzymatic activity levels in each population were compared with those obtained for the CAREC susceptible strain, and significant differences were determined by Kruskal-Wallis and Tukey's non-parametric tests (P<0.05). The established DDs were 0.01 mg l(-1), 0.2 mg l(-1) and 1.0 mg l(-1) for deltamethrin, permethrin and DDT, respectively; and the RTs for deltamethrin, permethrin and DDT were 30, 75 and 120 min, respectively. All Ae. aegypti populations were resistant to DDT (<80% mortality); two strains were incipiently resistant to deltamethrin and three to permethrin (80-98% mortality). Biochemical assays revealed elevated levels of α-esterase and MFO enzymes in all Ae. aegypti populations. All, except three populations, showed increased levels of β-esterases; and all populations, except Curepe, demonstrated elevated GST levels.Metabolic detoxification of enzymes is correlated with the manifestation of DDT and PY resistance in Trinidad and Tobago populations of Ae. aegypti. The presence of this resistance also suggests that knock down (kdr)-type resistance may be involved, hence the need for further investigations. This information can contribute to the development of an insecticide resistance surveillance programme and improvement of resistance management strategies aimed at combatting the spread of dengue in Trinidad and Tobago.     

Multiple insecticide resistance in Aedes aegypti (Diptera: Culicidae) populations compromises the effectiveness of dengue vector control in French Guiana

In French Guiana, pyrethroids and organophosphates have been used for many years against Aedes aegypti. We aimed to establish both the resistance level of Ae. aegypti and the ultra low volume spray efficacy to provide mosquito control services with practical information to implement vector control and resistance management. Resistance to deltamethrin and fenitrothion was observed. In addition, the profound loss of efficacy of AquaK'othrine? and the moderate loss of efficacy of Paluthion? 500 were recorded. Fenitrothion remained the most effective candidate for spatial application in French Guiana until its removal in December 2010. Further investigation of the mechanism of resistance to deltamethrin demonstrated the involvement of mixed-function oxidases and, to a lesser extent, of carboxylesterases. However, these observations alone cannot explain the level of insecticide resistance we observed during tube and cage tests.     

Evaluation of methoprene effect on Aedes aegypti (Diptera: Culicidae) development in laboratory conditions

Several Brazilian Aedes aegypti populations are resistant to the larvicidae temephos. Methoprene, that inhibits adult emergence, is one of the alternatives envisaged by the Brazilian Dengue Control Program (PNCD). However, at Brazil vector infestation rates are measured through larvae indexes and it has been claimed that methoprene use in the field could face operational problems. In order to define a standardized protocol, methoprene effect was evaluated in laboratory conditions after continuous exposure of larvae (Rockefeller strain) to a methoprene formulation available to the PNCD. Methoprene-derived mortality occurs mainly at the pupa stage and pupa development is inversely proportional to methoprene concentration. Number and viability of eggs laid by treated and control females are equivalent. A methoprene dose-dependent delay in the development was noted; however, strong correlations were found for total mortality or adult emergence inhibition if data obtained when all control mosquitoes have emerged are compared to data obtained when methoprene-treated groups finish development. The cumulative record of total methoprene-induced mortality at the time control adults emerge is proposed for routine evaluation of field populations. Mortality of all specimens, but not of larva, could account for adult emergence inhibition, confirming the inadequacy of larvae indexes to evaluate methoprene effect.     

Changes in enzyme titres with age in four geographical strains of Aedes aegypti and their association with insecticide resistance

Abstract. The enzymes acetylcholinesterase, glutathione S-transferase (GST), glucose 6-phosphate dehydrogenase (G6PD), and general esterases were assayed in four strains of Aedes aegypti mosquitoes aged between 1 and 30 days. Microtitre plate methods were used to assay activity in the homogenates of individual mosquitoes. The levels of GST and G6PD declined with the age of the mosquitoes, while the activity for the other enzymes remained constant. Soluble protein content was also found to decline with mosquito age in all the strains. Insecticide bioassays showed that two strains (Trinidad and Virtudes) of Ae. aegypti were resistant to DDT, deltamethrin and malathion, whereas two other strains (Bangkok and Indian) were susceptible to all four classes of insecticides tested. Higher esterase activity levels in the resistant compared to the susceptible strains were assumed to be the cause of organophosphate resistance. The combination of DDT and deltamethrin resistance in two strains with normal GST and G6PD characteristics suggests that a kdr-type nerve insensitivity mechanism may be involved.     

The efficacy of a chitin synthesis inhibitor against field populations of organophosphate-resistant Aedes aegypti in Brazil

The mosquito Aedes aegypti is the main focus of dengue control campaigns. Because of widespread resistance against conventional chemical insecticides, chitin synthesis inhibitors (CSIs) are considered control alternatives. We evaluated the resistance status of four Brazilian Ae. aegypti populations to both the organophosphate temephos and the pyrethroid deltamethrin, which are used in Brazil to control larvae and adults, respectively. All vector populations exhibited high levels of temephos resistance and varying rates of alterations in their susceptibility to pyrethroids. The effect of the CSI novaluron on these populations was also investigated. Novaluron was effective against all populations under laboratory conditions. Field-simulated assays with partial water replacement were conducted to evaluate novaluron persistence. Bioassays were continued until an adult emergence inhibition of at least 70% was attained. We found a residual effect of eight weeks under indoor conditions and novaluron persisted for five-six weeks in assays conducted in an external area. Our data show that novaluron is effective against the Ae. aegypti populations tested, regardless of their resistance to conventional chemical insecticides.     

The effects of Plagiorchis noblei metacercariae on the development and survival of Aedes aegypti larvae in the laboratory

Plagiorchis noblei infections impair the survival and development of fourth instar Aedes aegypti larvae. Mortality during the larval and pupal stages reached 92%, and 60% of the emerging adults were malformed. The metacercariae interfere with pupation and the emergence of adults. Larvae and pupae that fail to transform to the next developmental stage within the normal time characteristically persist for extended periods, but invariably die without transforming. Whereas 82% of the control larvae gave rise to functional adults, only 4% of infected larvae managed to do so. Such effects may facilitate the transmission of the parasite.     

Effect of four chlorophytes on larval survival, development and adult body size of the mosquito Aedes aegypti

The effect of four microalgal chlorophytes, Chlorella vulgaris,Scenedesmus quadricauda, Chlorococcum sp. and Ankistrodesmus convolutus, on the survival, larval development and adultbody size of the mosquito Aedes aegypti was investigated. The percentage mortality of larvae fed with C. vulgaris, Chlorococcum sp.or S. quadricauda after six days was 100%, 84% and 88%,respectively. Delayed pupation and body size reduction of the mosquitoesfed with C. vulgaris, Chlorococcum sp. and S. quadricaudawere observed. In contrast, larvae fed with A. convolutus werebigger than those fed with normal insectory feed. The study showed thatC. vulgaris, S. quadricauda and Chlorococcum sp. have potentialto be used as larvicidal agents.     

Oscillations of voltage and resistance in Malpighian tubules of Aedes aegypti

The transepithelial voltage (V(t)) of isolated Malpighian tubules of the yellow fever mosquito Aedes aegypti spontaneously oscillates in more than half the tubules. Typically, V(t) decreases and then rises at a frequency of 2 oscillations/min with a duration of 16 s. In 6 isolated perfused tubules studied in detail, V(t) oscillates between 50.5 mV and 15.7 mV in parallel with (1) oscillations of the transepithelial resistance (R(t)) between 7.61 kOmegacm and 3.63 kOmegacm, (2) oscillations of the basolateral membrane voltage of principal cells between -56.7 mV and -72.2 mV, and (3) oscillations of the apical membrane voltage between 107.2 mV and 87.8 mV. The oscillations are dependent on the Cl concentration in the extracellular solutions. As R(t) decreases during the oscillations V(t) goes to the transepithelial equilibrium potential of Cl (E(cl)) indicating transient changes in transepithelial Cl conductance as the mechanism of voltage and resistance oscillations. Since the largest voltage oscillations take place across the whole epithelium and not across cell membranes, oscillating Cl conductances are localized to a single transepithelial Cl diffusion barrier such as the paracellular pathway. This conclusion is supported by the analysis of electrically equivalent circuits that identify the shunt pathway as the site of oscillating Cl conductances.     

A laboratory investigation of mosquito larval predation by Toxorhynchites moctezuma on Aedes aegypti

1. Functional responses of predatory Toxorhynchites moctezuma (Dyar & Knab) larvae feeding on Aedes aegypti (L.) larvae (Diptera: Culicidae) were found to be type II of Holling (1959) and Rogers (1972). 2. Estimates of searching rate were generally higher for later instar predators. The search rate of second instar predators declined as prey instar increased, but fourth instar Tx. moctezuma had the highest search rate for second instar Ae. aegypti. 3. Prey handling times were higher for early instar predators and late instar prey. 4. When presented with mixtures of two instars of Ae. aegypti, second instar Tx. moctezuma showed frequency independent selectivity for the early instars, whereas fourth instar predators showed frequency independent selectivity for the late instars of Ae. aegypti. There was no evidence of frequency dependent predation. Preferences appeared to be transitive. 5. Extended random predator equations, using parameters derived from the functional responses, did not adequately describe the outcome of predation in the prey mixture experiment, even when the possibility of optimal switching behaviour was accounted for.     

Organization, ultrastructure, and development of midgut visceral muscle in larval Aedes aegypti

The midgut muscularis of larvae of the mosquito Aedes aegypti takes the form of a grid of longitudinal and circular muscle bands. The longitudinal and circular bands overlap at near right angles at many areas of intersection. The longitudinal bands run the length of the midgut. However, some bands of circular muscle, located in the anterior midgut, pass only partway around the gut. An unusual feature was observed at some regions where longitudinal and circular bands of muscle intersect: filaments oriented at near right angles to one another were present in the same membrane-bound fiber. These cruciform regions send contractile elements into both circular and longitudinal bands. The muscularis was fixed in a contracted state, so most of the sarcomeres are represented by complete overlap of myosin and lighter staining actin filaments. Features characteristic of supercontracting muscle, including perforated Z-lines, were seen in sarcomeres of circular muscle bands. Small invaginations resembling transverse tubules were present but a sarcoplasmic reticulum was not observed. While occasional cells that may be neurons or neurosecretory cells were observed, a network that might serve to coordinate the segmentation and peristaltic movement of the muscularis was not apparent.     

Effects of Toxorhynchites moctezuma larval predation on Aedes aegypti populations: experimental evaluation

Abstract. Predatory larvae of the mosquito Toxorhynchites moctezuma were used experimentally to control a standing crop of larvae of the dengue vector mosquito Ae.aegypti. Each week, fifty Ae.aegypti first instar larvae were introduced to each of five water-filled drums (220 litres) of the type commonly used for domestic water storage in Caribbean dwellings. At the beginning of the fourth week, a certain number (0, 1, 2, 5 or 10) of first instar Tx.moctezuma larvae were introduced to each drum and the daily yield of Ae.aegypti adults from each drum was monitored thereafter. The experiment was repeated three times. With only one or two Tx.moctezuma larvae, predation on Ae.aegypti larvae stopped the output of Ae.aegypti adults for 1 week. Five or ten Tx. moctezuma prevented any Ae.aegypti emergence for up to 16 weeks. Cannibalism among Tx. moctezuma larvae was seldom observed and appeared not to be a hindrance in using this species against Ae.aegypti. Thus Tx.moctezuma is regarded as a good candidate for the biological control of Ae.aegypti by augmentative releases.