A rapid method for screening mosquito repellents on Anopheles pseudopunctipennis and Aedes aegypti.

One of the main vectors for malaria in Latin America is Anopheles pseudopunctipennis (Theobald), whereas Aedes aegypti (L.) is the primary vector of dengue, yellow fever, Zika and chikungunya viruses. The use of repellents is recommended as a personal protection method against these mosquitoes. However there are very few studies evaluating the effect of repellents on An. pseudopunctipennis. The use of a Petri dish to study repellence has been applied by several authors on flies, cockroaches, kissing bugs and mosquitoes, being a valuable technique for species that are difficult to breed under laboratory conditions, such as An. pseudopunctipennis. In the present study, we evaluated the repellence of the essential oil of the Eucalyptus nitens (Shining gum), its main component (1,8-cineole) and the commercial repellent DEET on An. pseudopunctipennis and Ae. aegypti adult females using the plaque repellency method coupled to EthoVision XT10.1 video-tracking software. Repellent effect and locomotor activity were studied through a repellence index (RI) together with an axis constructed from the behavioural variables obtained using the tracking software. DEET repellent effect was observed at 0.01 mg/mL for Ae. aegypti and 0.01 and 0.1 mg/mL for An. pseudopunctipennis. In addition, the essential oil showed significant repellence at 1 and 10 mg/mL for Ae. aegypti, and 1, 5, 10 and 25 mg/mL for An. pseudopunctipennis. Neither of these species were repelled at any concentration of 1,8-cineole. This is the first study that evaluates these compounds on An. pseudopunctipennis females and quantifies their effects on the activity of both species.

First Report of Adult Aedes aegypti (Diptera: Culicidae) Resistance to Pyrethroids in Argentina.

Severe human arboviral diseases can be transmitted by the mosquito Aedes aegypti (Linnaeus), including dengue, chikungunya, Zika, and yellow fever. Adult control using spatial sprays with adulticides is recommended only when dengue outbreaks occur. In Argentina, mainly pyrethroids, like cis-permethrin, have been used as an adulticide, especially since 2008. The evolution and spread of resistance to insecticides is a major concern for vector control. This study reports for the first time pyrethroid resistance in Ae. aegypti adults from Argentina, in the city of Salvador Mazza (Salta). WHO discriminating doses of 0.75% were used for permethrin, 0.05% for deltamethrin, and 5% for malathion. Also the discriminating dose for cis-permethrin (0.6%) was calculated and evaluated for the first time. We found a resistance ratio 50 (RR50) of 10.3 (9.7-10.4) for cis-permethrin, which is considered as high resistance. Our results also indicated resistance to deltamethrin (22.6% mortality) and permethrin (53.6% mortality), and a total susceptibility to malathion (100% mortality). Results from this study highlight the importance of the correct use of insecticides within an Integrated Vector Management (IVM) approach and of early detection of resistance to enable Ae. aegypti control in Argentina. More studies are needed to determine the spread of mosquito resistance to pyrethroids.

Lufenuron can be transferred by gravid Aedes aegypti females to breeding sites and can affect their fertility, fecundity and blood intake capacity.

BACKGROUND: Aedes aegypti (L.) is the main vector of dengue, yellow fever, Zika and chikungunya viruses. A new method for controlling this mosquito has been developed based on the possibility that wild adult mosquitoes exposed to artificial resting sites contaminated with a larvicide, can disseminate it to larval breeding sites, is named "auto-dissemination". The present study was undertaken to evaluate if a chitin synthesis inhibitor like lufenuron can be disseminated to larval breeding sites and prevent adult emergence and also if forced contact of Ae. aegypti females with treated surfaces can affect its fertility, fecundity, and blood intake capacity. METHODS: Larval susceptibility to lufenuron was measured through EI50 and EI90. On the other hand, gravid females were exposed by tarsal contact to lufenuron-treated papers, we used the WHO susceptibility test kit tube to line the papers, and 1, 3 or 5 females for the transference. We also evaluated if the exposure of female mosquitoes to lufenuron-treated papers (0.4 and 1 mg a.i./cm2) has an effect on their fertility, fecundity or in the ability to feed on blood. In each assay 12-15 female mosquitoes were exposed to lufenuron for 1 h, 24 h before blood meal (BBM) or 24 h after a blood meal (ABM). RESULTS: Lufenuron proved to be very active against Ae. aegypti larvae with an EI50 of 0.164 ppb and EI90 of 0.81 ppb. We also found that lufenuron can be transferred by females from treated surfaces to clean containers causing the inhibition of emergence of the larvae (between 30 and 50%). This effect was dependent on the concentration applied on the paper and the number of females added to each cage. CONCLUSIONS: This study introduces an innovation by first exploring the possibility that an insect growth regulator (IGR) belonging to the group of benzoylphenyl ureas, such as lufenuron, can be transferred by gravid females to breeding sites and that at the same time can have an effect on fertility, fecundity and blood intake capacity of adult mosquitoes.

Soil presence reduces the control effectiveness of a slow-release formulation of pyriproxyfen on Aedes aegypti (Diptera: Culicidae) larvae.

OBJECTIVE: To assess the influence of soil on the effectiveness of two new slow-release formulations (floating and non-floating) of pyriproxyfen coextruded with low-density polyethylene. METHODS: Two slow-release devices were developed using low-density polyethylene, pyriproxyfen as larvicide and calcium carbonate as filler. A factorial design was used to evaluate the effect of soil presence on the performance of each device. Weekly bioassays were performed. RESULTS: Soil presence affected treatment effectiveness, but this effect was associated with device type. The tablets were effective for nearly 3 months. CONCLUSION: Treatment effectiveness could be reduced because of the loss of pyriproxyfen by several physico-chemical processes such as adsorption into the soil.

OBJECTIF: Evaluer l'influence de la terre sur l'efficacité de deux nouvelles formulations à libération lente (flottante et non flottante) de pyriproxyfène coextrudé avec du polyéthylène de faible densité. MÉTHODES: Deux dispositifs à libération lente ont été développés en utilisant du polyéthylène de faible densité, du pyriproxyfène comme larvicide et du carbonate de calcium comme agent de charge. Un plan factoriel a été utilisé pour évaluer l'effet de la présence de terre sur les performances de chaque dispositif. Des essais biologiques hebdomadaires ont été effectués. RÉSULTATS: La présence de terre a affecté l'efficacité du traitement, mais cet effet était associé au type de dispositif. Les comprimés ont été efficaces pendant près de 3 mois. CONCLUSION: L'efficacité du traitement pourrait être réduite en raison de la perte du pyriproxyfène par plusieurs processus physicochimiques tels que l'adsorption sur la terre.

Effects of Temephos, Permethrin, and Eucalyptus nitens Essential Oil on Survival and Swimming Behavior of Aedes aegypti and Anopheles pseudopunctipennis (Diptera: Culicidae) Larvae.

An essential strategy to deal with mosquito-borne diseases is the control of larvae in their development sites. The mosquitoes Anopheles pseudopunctipennis (Theobald) (Diptera: Culicidae), a malaria vector, and Aedes aegypti (L.) (Diptera: Culicidae), vector of dengue, Zika, yellow fever, and chikungunya viruses, breed in very different habitats. Insecticide treatments of mosquito larvae focus mainly on their lethal effects. However, insecticide degradation or the poor dosage of larvicides will invariably lead to the sublethal exposure of a target (and nontarget) species, the nonlethal effects of these compounds may have important effects on vital insect activities, and therefore their evaluation is necessary. In this study, we assessed the survival and swimming behavior of larvae of Ae. aegypti and An. pseudopunctipennis exposed to increasing concentrations of three larvicides. We found that Ae. aegypti, was more sensitive to the larvicides than An. pseudopunctipennis, we also observed an overall decrease in the movement of those larvae of both species, which survive the treatments. This decrease might have ecological relevance in their natural habitats, increasing the chance to be predated and decreasing their ability to obtain food. Finally, this information will be valuable to assist authorities to make decisions in the implementation of further control programs.

N-substituted methyl maleamates as larvicidal compounds against Aedes aegypti (Diptera: Culicidae).

Severe human arboviral diseases can be transmitted by the mosquito Aedes aegypti (L.), including dengue, chikungunya, zika, and yellow fever. The use of larvicides in containers that can result as potential breeding places and cannot be eliminated is the main alternative in control programs. However, their continuous and widespread use caused an increase in insecticide-resistant populations of this mosquito. The aim of this study was to evaluate the effect of three N-substituted methyl maleamates as larvicides on Ae. aegypti, the N-propyl methyl maleamate (PMM), N-butyl methyl maleamate (BMM), and N-hexyl methyl maleamate (HMM). These compounds could have a different mode of action from those larvicides known so far. We evaluated the larva mortality after 1 and 24 h of exposure and we found that mortality was fast and occurs within the first 60 min. HMM was slightly more effective with LC50 values of 0.7 and 0.3 ppm for 1 and 24 h of exposure and LC95 of 11 and 3 ppm. Our results demonstrate that N-substituted methyl maleamates have insecticidal properties for the control of Ae. aegypti larvae. These compounds could become useful alternatives to traditional larvicides after studying their insecticidal mechanism as well as their toxicity towards non target organisms.

Biological and Chemical Characterization of a New Larvicide Ovitrap Made of Plastic With Pyriproxyfen Incorporated for Aedes aegypti (Diptera: Culicidae) Control.

Aedes aegypti (L.) is a species of international concern because of its ability to transmit serious human arboviral diseases including yellow fever, dengue, and chikungunya, which have spread to all continents. Ovitraps are containers constructed to imitate Aedes' natural breeding sites and have been used for many decades as a sensitive and inexpensive surveillance tool for detecting the presence of container-inhabiting mosquitoes. In addition to their value for vector surveillance, various ovitrap devices have been evaluated as tools for suppressing Ae. aegypti populations. In this study, we performed a biological and chemical characterization of a new ovitrap prototype manufactured by injection molding of low-density polyethylene (LDPE) with the larvicide pyriproxyfen. Our research shows that pyriproxyfen was immediately released from the LDPE into the water of the ovitrap and led to an emergence inhibition of 100% for over 30 weeks. In addition, ovitraps continued to show a high larvicidal activity after over 20 washes. Pyriproxyfen was detectable in the water after 20 s and reached a peak after 24 h. Our results show that this ovitrap can be an effective, inexpensive, and low-maintenance tool for Ae. aegypti surveillance and control.

The use of Aedes aegypti larvae attractants to enhance the effectiveness of larvicides.

Aedes aegypti (L.) is an important dengue, chikungunya, and yellow fever vector. Immature stages of this species inhabit human-made containers placed in residential landscapes, and the application of larvicides inside containers that cannot be eliminated is still considered a priority in control programs. Larvicidal efficacy is influenced by several factors, including the formulation used, the water quality, and the susceptibility of larvae, among others. If an attractant can be incorporated into a slow-release larvicide formulation, it will be feasible to direct the larvae into the source of insecticide and thereby improving its efficacy. We studied the influence of 1-octen-3ol and 3-methylphenol on the rate of Ae. aegypti larvae mortality using the larvicides Bacillus thuringiensis var. israelensis (Bti), temephos, and spinosad. These chemicals were combined with the larvicides mixed with agar during the bioassays. Mortality was registered every 10 min, and a lethal time 50 (LT50) was calculated. The inclusion of the Ae. aegypti larvae attractants with the larvicides into a solid agar matrix improved their efficiency obtaining a strong and marked reduction in the LT50 compared with the use of larvicides alone.

Sublethal effect of pyriproxyfen released from a fumigant formulation on fecundity, fertility, and ovicidal action in Aedes aegypti (Diptera: Culicidae).

Dengue and dengue hemorrhagic fever are mosquito-borne viral diseases that coincide with the distribution of Aedes aegypti (L.), the primary vector in the tropical and semitropical world. With no available vaccine, controlling the dengue vector is essential to prevent epidemics. The effects of the insect growth regulator pyriproxyfen on Ae. aegypti adults that survived a treatment with a sublethal dose were investigated in the laboratory, including effects on their reproductive potential. Pyriproxyfen was released from a fumigant formulation at a dose causing 20 or 40% emergence inhibition (%EI). Females were dissected before and after blood feeding and the basal follicle number was counted. There were no differences between the control and treated group on the basal follicle number for both doses used. Fertility and fecundity were reduced at a concentration of EI40 but no at EI20. There was no ovicidal effect of pyriproxyfen by immersion of eggs in treated water neither when the females laid their eggs on a pyriproxyfen-treated surface. This work shows that sublethal doses of pyriproxyfen can have effects on fertility and fecundity ofAe. aegypti females, which together with its larvicidal activity could contribute to an overall decrease in a given population.

Field comparison of thermal and non-thermal ultra-low-volume applications using water and diesel as solvents for managing dengue vector, Aedes aegypti.

OBJECTIVES: To compare the effectiveness on Aedes aegypti (Linneo) (Diptera: Culicidae) of a larvicide-adulticide ULV formulation applied by a thermal or a cold fogger using different solvents. METHODS: We applied, in field conditions, a ULV formulation containing pyriproxyfen and permethrin, using a thermal and a cold fogger and water or diesel as solvent. We determined the effectiveness of these applications on Ae. aegypti adults and larvae by different bioassays and measuring Breteau, house and adult indices. RESULTS: When water was used as solvent, the treatments applied with the cold or the thermal foggers were equally effective on adult mortality (close to 90%) and adult emergence inhibition (% EI) (close to 70%). When the thermal fogger was used with water as solvent, the adult mortality outside the houses (85%) was higher, but not significantly different, than with diesel (65%). The contrary happens inside (22%vs. 58%), while there were no differences in %EI. Adult and larval indices behaved similarly in all areas, with a slight tendency for the treatments applied using water as solvent to be more effective. CONCLUSIONS: Water-based formulations are equally or more effective than the one applied with diesel as solvent. The use of water as solvent will not only improve the effectiveness of this formulation but also reduce the environmental impact and costs of spraying compared to the use of diesel.

Droplet size and efficacy of an adulticide-larvicide ultralow-volume formulation on Aedes aegypti using different solvents and spray application methods.

BACKGROUND: When cases of dengue are reported or the density of adult Aedes aegypti (L.) becomes too high, ultralow-volume (ULV) application of insecticides is the recommended control method. The droplet size of an aerosol insecticide influences its efficiency in killing adult mosquitoes. Many studies have been carried out to determine the optimum droplet size that maximises vector control efforts, but only a few have determined droplet-size spectra for specific equipment using different solvents and comparing thermal and non-thermal aerosols. RESULTS: The present study showed that the droplet size for a water-based adulticide-larvicide formulation was larger than for the same formulation diluted in gasoil or biodiesel. No significant differences in adult mortality were observed between sprayers and solvents, but efficacy decreased with distance from the sprayer nozzle. Adult emergence inhibition was more than 90% when using water as a solvent for both thermal and cold foggers, and the efficacy did not decrease with distance from the sprayer nozzle. On the other hand, oil-based solvents became less effective with distance. CONCLUSION: The use of water as a solvent with both thermal and cold foggers improves the efficacy of the studied formulation containing permethrin as adulticide and pyriproxyfen as larvicide in scaled-up assays. Moreover, it reduces the environmental impact and costs of spraying by comparison with formulations using oil solvents.

A new strategy for Aedes aegypti (Diptera: Culicidae) control with community participation using a new fumigant formulation.

Dengue and dengue hemorrhagic fever are mosquito-borne viral diseases that coincide with the distribution of Aedes aegypti (L.), the primary vector in the tropical and semitropical world. With no available vaccine, controlling the dengue vector is essential to avoid epidemics. This study evaluates the efficacy of a new smoke-generating formulation containing pyriproxyfen and permethrin in Puerto Libertad, Misiones, Argentina. A fumigant tablet (FT) was applied inside the houses by the community members and compared with a professional application. A treatment combining the application of fumigant tablets indoors and ultralow volume fumigation outdoors was also assessed. The community perceptions and practices about dengue disease and the acceptance of this new nonprofessional FT were evaluated through surveys. Results show >90% adult emergence inhibition and 100% adult mortality with these treatments. More than 80% of the residents applied the FT and preferred participating in a vector control program by using a nonprofessional mosquito control tool, instead of attending meetings and workshops promoting cultural changes.

Thermal behaviour and biological activity against Aedes aegypti (Diptera: Culicidae) of permethrin and pyriproxyfen in a smoke-generating formulation.

BACKGROUND: The most common ways to control dengue vector Aedes aegypti (L.) are larval source reduction in domestic habitats and ground application of small quantities of aerosol insecticide (ultralow volume). Nevertheless, these actions have been shown repeatedly to be ineffective in controlling Ae. aegypti populations. RESULTS: The efficacy of a new smoke-generating formulation containing pyriproxyfen and permethrin was evaluated in the laboratory. Smoke-generating tablets containing each insecticide individually or combined were prepared, and the recovery of the insecticides from the smoke was determined. Recovery values of over 90% were obtained for pyriproxyfen, and around 50% for permethrin. The biological efficacy of pyriproxyfen released in the smoke was evaluated in the laboratory, on late third-instar or early fourth-instar Ae. aegypti larvae, using different concentrations of pyriproxyfen and exposure times. Adult emergence inhibition (EI) values of 100% were obtained at 30 min, and a dose-dependent effect was observed at 5 min. The effect of pyriproxyfen released in the smoke was due to direct contact with the larvicide in the water rather than by inhalation of the fumes. The efficacy of permethrin released in the fumes was also evaluated as knockdown effect (KT(50)) on adults for a tablet containing permethrin alone or permethrin plus pyriproxyfen. There was no significant difference in KT(50) values obtained for permethrin (KT(50) = 19.9 min) and permethrin plus pyriproxyfen (KT(50) = 19.4 min). CONCLUSION: The excellent laboratory performance of this new formulation on immature stages and adults indicates that a smoke-generating tablet containing pyriproxyfen and permethrin could be a new tool for controlling mosquitoes.

Efficacy of a new combined larvicidal-adulticidal ultralow volume formulation against Aedes aegypti (Diptera: Culicidae), vector of dengue.

A new ultralow volume formulation (ULV) containing permethrin as an adulticidal active ingredient and the insect growth regulator (IGR) pyriproxyfen as a larvicide was developed and its efficacy evaluated in a field trial in Wanda, Misiones (Argentina). Two separate study areas were sprayed: one with a ULV formulation of permethrin 15% plus pyriproxyfen 3% and the other with permethrin 15% only. A third untreated area was kept as a control. Sentinel cages containing adult mosquitoes and jars containing Aedes aegypti larvae III/IV were placed in treated and control areas. The residual activity of the formulations was tested using 20 L water containers. The adulticide effect of permethrin 15% + pyriproxyfen 3% formulation was similar to the permethrin 15% formulation, however, the inhibition of adult emergence in the treatment with permethrin 15% never exceeded 20%, whereas the inhibition of adult emergence in the treatment with permethrin 15% plus pyriproxyfen 3% showed initial values of 96% maintaining this high level of inhibition up to 35 days after ULV spraying. Larval indexes (House and Breteau indexes) showed that a greater, long-lasting effect was obtained with the permethrin 15% plus pyriproxyfen 3% formulation.

Effectiveness of pyriproxyfen and diflubenzuron formulations as larvicides against Aedes aegypti.

The efficacy of technical pyriproxyfen and diflubenzuron was evaluated at the laboratory level against Aedes aegypti, where we achieved adult emergence inhibition (EI50) values of 0.048 and 1.59 ppb (microg/liter), respectively. We compared these data with values obtained for temephos. We prepared emulsifiable concentrate formulations of pyriproxyfen and diflubenzuron in the laboratory and obtained EI50 values of 0.01 and 0.02 ppb, respectively. We also obtained 100% adult emergence inhibition with 0.1% slow-release sand formulations of pyriproxyfen and diflubenzuron. In a simulated field study, the granular sand formulations of pyriproxyfen, diflubenzuron, and temephos remained active for over 4 mo. In a field study, a 0.2% granular formulation of diflubenzuron and 1% temephos showed a similar performance.