Two Lactones Non-toxic to Humans Are Effective Repellents for the German Cockroach.

Lactones are cyclic esters of hydroxy carboxylic acids, present in several fruits and animals consumed by humans. There is evidence that some lactones modify insect behavior. The aim of the present study was to evaluate the repellent effect of four lactones (γ- and δ-nonalactone, and γ- and δ-dodecalactone) in first instar nymphs of the German cockroach, Blattella germanica (Linnaeus). To assess repellency, a nymph was placed on a circle of filter paper, half of which had been treated with lactone dissolved in acetone and the other half with acetone alone. The behavior of the nymph was recorded and the time the nymph spent in each half of the paper was quantified using Ethovision XT 10.1 software. Values of Distribution Coefficient (DC) were calculated: DC = (Tt - TA) / Tt, where Tt is the experimental time and TA is the time the nymph spent in the area treated with the repellent agent. DC can vary between 0 and 1. Values significantly higher than 0.5 indicate repellency. N,N-diethyl-meta-toluamide (DEET) was used as a positive control. DEET, δ- and γ-nonalactone caused repellency as from 77.9 µg/cm2, whereas γ- and δ-dodecalactone had a repellent effect starting at 779.0 µg/cm2. The values of DC for these concentrations were 0.89 (DEET), 0.86 (γ-nonalactone), 0.87 (δ-nonalactone), 0.83 (γ-dodecalactone), and 0.72 (δ-dodecalactone). To our knowledge, this is the first report of repellency produced by lactones in the German cockroach. This work allowed to identify two lactones that have a repellent effect similar to DEET.

Eugenol and menthol synergize the toxicity of permethrin in the blood-sucking bug, Triatoma infestans.

The blood-sucking bug, Triatoma infestans, is one of the main vectors of Chagas disease in America. It is usually controlled with pyrethroids, but the emergence of resistance to these insecticides creates the need to look for alternative products. Eugenol, menthol and menthyl acetate are botanical monoterpenes, which produce lethal and sublethal effects on insects. The purpose of this work was to determine what type of toxicological interactions occur when binary mixtures, formed by the pyrethroid permethrin and sublehtal doses of eugenol, menthol or menthyl acetate, are applied to T. infestans. First instar nymphs were exposed to filter papers impregnated with the insecticides. The number of knocked down insects was registered at different times and Knock Down Time 50% (KT50) values were calculated. The following KT50 values with their corresponding 95% Confidence Intervals were obtained: permethrin, 47.29 (39.92 - 56.32) min; permethrin + eugenol, 34.08 (29.60 - 39.01) min; permethrin + menthol, 27.54 (23.28 - 32.55) min; permethrin + menthyl acetate, 43.62 (39.99 - 47.59) min. Eugenol and menthol increased the speed of action of permethrin (synergism), but menthyl acetate had no effect on it (additivity). These results provide the basis to further explore interactions between conventional insecticides and plant monoterpenes as potential tools for controlling T. infestans.

Fitness and mating compatibility of Chrysodeixis includens (W.) (Lepidoptera: Noctuidae) populations collected in different provinces and crops in Argentina.

Chrysodeixis includens, is an important Plusiinae pest that feeds on soybean, beans, and cotton. This species has a genetic structure determined by molecular markers in populations collected in soybean crops in Brazil and has developed resistance to insecticides. This had led to an increased interest in the genetic diversity of its populations in Argentina. This study aimed to characterize the fitness parameters and mating compatibility of geographically distant C. includens populations and on different host plants in Argentina. Intra- and inter-population crosses revealed that C. includens populations from Tucumán and Santiago del Estero provinces did not present any evidence of prezygotic and postzygotic incompatibilities, suggesting that both populations collected from soybean plants belonged to a single wide-ranging species. Prezygotic and postzygotic incompatibilities were detected between populations collected from different crops (soybean and bean). These incompatibilities may be caused by the different nutrients that each plant species renders, or the specific pest control strategies which may affect C. includens behavior. Studies of the genetic structure of C. includens populations based on mating isolation on other crops, as well as of the impact of management strategies on the specie, will increase understanding of the reproductive compatibility revealed by the present study.

The Octopamine Receptor Is a Possible Target for Eugenol-Induced Hyperactivity in the Blood-Sucking Bug Triatoma infestans (Hemiptera: Reduviidae).

Eugenol is a major component of the essential oils in cloves and other aromatic plants. In insects, it produces toxic effects and repellency, and there is evidence that its site of action is the octopamine receptor. The objective of the present study was to explore whether the octopamine receptor is involved in the hyperactivity produced by eugenol in the blood-sucking bug Triatoma infestans (Klug). This insect is the main vector of Chagas disease in Latin America. Four treatments were topically applied on third instar nymphs: 1) octopamine, 2) eugenol, 3) phentolamine hydrochloride (an antagonist of the octopamine receptor) followed by octopamine, and 4) phentolamine hydrochloride followed by eugenol. Both octopamine and eugenol hyperactivated the nymphs. However, pretreatment with phentolamine hydrochloride inhibited the hyperactivating effect of both compounds. These results are in agreement with previous works on Drosophila melanogaster (Meigen) (Diptera: Drosophilidae) and the American cockroach. They suggest that the octopamine receptor is a possible site of action for eugenol.

Deltamethrin-resistant German Cockroaches Are Less Sensitive to the Insect Repellents DEET and IR3535 than Non-resistant Individuals.

The German cockroach, Blattella germanica (L.) (Blattodea: Blattellidae), is a serious worldwide pest with a considerable economical and sanitary impact. It is mainly controlled by the application of synthetic insecticides, but repeated use of these substances has promoted the appearance of resistance in cockroach populations throughout the world. The aim of this study was to compare the behavior of deltamethrin-susceptible (CIPEIN colony) and deltamethrin-resistant (JUBA and VGBA colonies) first instar nymphs exposed to the repellents N,N-diethyl-3-methylbenzamide (DEET) and ethyl 3-[acetyl(butyl)amino]propanoate (IR3535). Firstly, the behavior of the nymphs was assessed in an experimental arena in the absence of repellents. The parameters Distance Traveled, Velocity, Mobility Time, and Time Spent (in each half of the arena) were quantified using an image analyser, and showed that the behavior elicited by the three colonies was similar. After this, the behavior of the nymphs was quantified in an arena, half of which had been treated with repellent. The repellency of DEET increased as a linear function of log concentration for the three colonies. DEET elicited repellency as from a concentration of 97.49 µg/cm2 for the CIPEIN and JUBA colonies and 194.98 µg/cm2 for the VGBA colony. The repellency of IR3535 was weaker and started at a concentration of 389.96 µg/cm2 for the CIPEIN colony, 779.92 µg/cm2 for JUBA, and 1559.84 µg/cm2 for VGBA . Finally, nymphs were exposed to 3:1, 1:1, and 1:3 DEET:IR3535 mixtures, and a synergistic effect was observed only in the CIPEIN colony.

Toxicity, repellency and flushing out in Triatoma infestans (Hemiptera: Reduviidae) exposed to the repellents DEET and IR3535.

DEET and IR3535 are insect repellents present worldwide in commercial products; their efficacy has been mainly evaluated in mosquitoes. This study compares the toxicological effects and the behavioral responses induced by both repellents on the blood-sucking bug Triatoma infestans Klug (Hemiptera: Reduviidae), one of the main vectors of Chagas disease. When applied topically, the Median Lethal Dose (72 h) for DEET was 220.8 µg/insect. Using IR3535, topical application of 500 µg/insect killed no nymphs. The minimum concentration that produced repellency was the same for both compounds: 1,15 µg/cm2. The effect of a mixture DEET:IR3535 1:1 was similar to that of their pure components. Flushing out was assessed in a chamber with a shelter containing groups of ten nymphs. The repellents were aerosolized on the shelter and the number of insects leaving it was recorded for 60 min. During that time, 0.006 g/m3 of the positive control tetramethrin flushed out 76.7% of the nymphs, while 1.76 g/m3 of DEET or IR3535 flushed out 30 and 0%, respectively. The concentrations required for both compounds to produce toxicity or flushing out are too high to have any practical applications. However, they showed a promising repellency. Additional research should be done to evaluate their possible use for personal protection against T. infestans bites.

Rhodnius prolixus intoxicated.

Rhodnius prolixus (Hemiptera: Reduviidae) is a hematophagous insect native from South America. By the end of the 20th century, it was one of the main vectors of Chagas disease in Venezuela, Colombia, several Central American countries and southern Mexico. The aim of the present article is to review the literature regarding R. prolixus toxicology. British entomologist Vincent B. Wigglesworth carried out the first studies on this subject over seventy years ago. A wide bibliographical search allowed to locate one hundred and thirty scientific articles describing the effects of different insecticides on R. prolixus. About one-third of these articles report the acute toxicity and/or sublethal effects produced by the main synthetic neurotoxic families of insecticides (organochlorines, organophosphates, carbamates and pyrethroids). Only a couple of these studies have regarded the toxicokinetics or toxicodynamics of these insecticides. Insect growth or development disruptors, such as juvenoids, chitin synthesis inhibitors, precocenes, azadirachtin and lignoids, have been thoroughly studied in R. prolixus. Important aspects on the mode of action of ureases were also described in this species. By the end of the 1960's, resistance to insecticides was detected in R. prolixus from Venezuela. Some years later, the existence of pyrethroid-resistant individuals was also reported. Control programmes for R. prolixus in countries where Chagas is endemic have only used synthetic neurotoxic insecticides. In 2011, Central America and southern Mexico were declared free of this insect. The recent sequencing of the R. prolixus genome will provide valuable information to understand the molecular basis of insecticide resistance in this species.

Field Evaluation of a New Strategy to Control Lutzomyia longipalpis, Based on Simultaneous Application of an Adulticide-Larvicide Mixture.

Leishmania infantum (syn. chagasi) is the etiologic agent of visceral leishmaniasis in Argentina, and the phlebotomine fly Lutzomyia longipalpis is its main vector. The objective of this study was to evaluate the effectiveness of Dragon Max®, an emulsifiable concentrate formulation containing the pyrethroid permethrin and the larvicide pyriproxyfen, for Lu. longipalpis control under field conditions. The work was conducted in the city of Posadas (Misiones province, Argentina). Comparisons were performed between treated and untreated peridomiciles with poultry, which met previously determined criteria for favoring the presence of Lu. longipalpis. Henhouses and their surrounding area were treated, with the formulation (100 mg of permethrin and 2 mg/m2 of pyriproxyfen) applied using a hand pump sprayer. Untreated henhouses were used as controls. Phlebotomine abundance was monitored before treatment and then weekly, using Centers for Disease Control and Prevention light traps. Lutzomyia longipalpis was the only phlebotomine species captured. A male/female ratio of 2.5 was observed. The more chickens there were in the henhouses, the greater the number of phlebotomines captured. The treatment resulted in a significant decrease in the number of individuals, which persisted for at least 2 wk. This encouraging result provides a baseline for further studies evaluating the possibility of using Dragon Max as a tool for Lu. longipalpis control.

Behavioral and Toxicological Responses of Rhodnius prolixus (Hemiptera: Reduviidae) to the Insect Repellents DEET and IR3535.

,N-diethyl-3-methylbenzamide (DEET) is a broad-spectrum insect repellent used by millions of people since the 1950s. Ethyl 3-[acetyl(butyl)amino]propanoate (IR3535) is a repellent developed more recently that is still not used as extensively. This study compares the behavioral and toxicological effects of both substances in fifth-instar nymphs of the blood-sucking bug Rhodnius prolixus Stål (Hemiptera: Reduviidae), one of the main vectors of American trypanosomiasis (Chagas disease). Repellency was quantified using filter paper discs as experimental arenas. Half the discs were treated with solution of repellent in acetone, and the other half with acetone alone. The lowest observed effect level was identical for both substances, 74 µg/cm2. Nymph age (between 1-3 h and 99 d from last ecdysis) had no influence on repellency. Topical application of 750 µg of DEET per nymph produced a mortality rate between 0% (24 h after application) and 40% (7 d later). The same dose of IR3535 produced no mortality during the same period of time. Simultaneous treatment with piperonylbutoxide (a mixed function microsomal oxidase inhibitor) synergized the lethal effect of DEET. Only DEET increased locomotor activity in nymphs exposed to a treated surface. Nymph antennectomy abolished DEET repellency but not its effect on locomotor activity. The concentrations of both these compounds required to produce either behavioral or toxicological effects are too high to have any practical applications in the control of R. prolixus.

Toxic effect of citrus peel constituents on Anastrepha fraterculus Wiedemann and Ceratitis capitata Wiedemann immature stages.

The toxicity of essential oils from the citrus peel has been proposed as the major resistance mechanism offered by citrus to fruit fly infestation. We evaluated the insecticidal activity of the ether extracts from the lemon (Citrus limon [L.] Burm.) and grapefruit (C. paradisi Macfadyen) peel as well as from limonene and citral against Anastrepha fraterculus (Wiedemann) and Ceratitis capitata (Wiedemann) immature stages. We also evaluated the toxicity of the extracts at two ripening stages. Extracts proved toxic to A. fraterculus egg and larvae. The lemon and grapefruit extracts showed the same toxicity in both fruit fly species. For A. fraterculus eggs, citral was more toxic than limonene; for larvae, they showed equal toxicity. Anastrepha fraterculus eggs were more sensitive than C. capitata eggs. In conclusion, we provide evidence of chemical resistance mechanisms that could account for the nonhost condition of lemon for A. fraterculus.

Locomotor behaviour of Blattella germanica modified by DEET.

N,N-diethyl-3-methylbenzamide (DEET) is the active principle of most insect repellents used worldwide. However, its toxicity on insects has not been widely studied. The aim of this work is to study the effects of DEET on the locomotor activity of Blattella germanica. DEET has a dose-dependent repellent activity on B. germanica. Locomotor activity was significantly lower when insects were pre-exposed to 700 µg/cm(2) of DEET for 20 or 30 minutes, but it did not change when pre-exposure was shorter. Locomotor activity of insects that were pre-exposed to 2.000 µg/cm(2) of DEET for 10 minutes was significantly lower than the movement registered in controls. No differences were observed when insects were pre-exposed to lower concentrations of DEET. A 30-minute pre-exposure to 700 µg/cm(2) of DEET caused a significant decrease in locomotor activity. Movement was totally recovered 24 h later. The locomotor activity measured during the exposure to different concentrations of DEET remained unchanged. Insects with decreased locomotor activity were repelled to the same extent than control insects by the same concentration of DEET. We demonstrated that the repellency and modification of locomotor activity elicited by DEET are non-associated phenomena. We also suggested that the reduction in locomotor activity indicates toxicity of DEET, probably to insect nervous system.

Behavioral and toxicological responses of Rhodnius prolixus and Triatoma infestans (Hemiptera: Reduviidae) to 10 monoterpene alcohols.

The effect on locomotor activity, the repellency, and the knock-down produced by 10 monoterpene alcohols were evaluated on first-instar nymphs of Rhodnius prolixus and Triatoma infestans, vectors of Chagas disease. A video tracking technique was used to evaluate locomotor activity and repellency by exposure to papers impregnated with monoterpenes. Eugenol on R. prolixus and (S)-cis-verbenol on T. infestans did not modify the locomotor activity. The remaining monoterpenes produced hyperactivity on both species, although the concentration required was at least a 1,000 times higher than that of deltamethrin (positive control). Carvacrol, eugenol, and geraniol resulted as repellent as N,N-diethyl-m-toluamide (positive control) for both species. A similar result was observed for almost every monoterpene on T. infestans. Knock-down effect was evaluated by exposing the nymphs in closed recipients. The order of increasing toxicity on R. prolixus was (KT50 values in min): geraniol (213.7) < alpha-terpineol (164.5) < linalool (124.2) < carvacrol (111.6) < eugenol (89.8) < thymol (78.9), and on T. infestans: alpha-terpineol (289.8) < eugenol (221.3) < carvacrol (164.2) < linalool (154.9) < thymol (96.7). All monoterpenes were less toxic than the positive control, dichlorvos (3.6 min for R. prolixus and 3.9 min for T. infestans). After 7 h of exposure, (-)-carveol, citronellol, and menthol (on both species) and geraniol (on T. infestans) produced < 50% of knock-down. After these results, it is worthwhile to explore more deeply the potential of these compounds as tools for controlling Chagas disease vectors.

Triatomicidal effect of new spot-on formulations applied to poultry in semi-field conditions.

Chagas disease is an endemic disease affecting ten million people in the American continent. Produced by a parasite transmitted by triatomine insects, the main actions for reducing the incidence of this disease are focused on the control of insect vectors. This type of control has produced highly effective results within rural homes, but not in peridomestic areas (kitchens, warehouses, hen houses and other buildings not attached to the houses). The object of the present study was to assess the triatomicidal effect of new spot-on formulations developed by our laboratory in a semi-rural environment. The active ingredients of the formulations were ß-cypermethrin, pyriproxyfen, or ß-cypermethrin + pyriproxyfen. All formulations were applied to hens and tested in miniature replicas of rural households where experimental populations of Triatoma infestans, the main vector of Chagas disease in Argentina, had been previously released. The experimental populations exposed to formulations containing ß-cypermethrin or ß-cypermethrin + pyriproxyfen were noticeably reduced compared to non-treated control groups. However, no differences were observed between the effects produced by ß-cypermethrin alone and ß-cypermethrin + pyriproxyfen. Pyriproxyfen alone produced no significant reduction in the experimental populations of T. infestans. These results suggest that spot-on application of ß-cypermethrin could be a useful complementary tool for controlling triatomine insects in the peridomestic areas of rural homes.

Adaptation of the repellency response to DEET in Rhodnius prolixus.

For many years it has been accepted that DEET interferes with the detection of odours from the host instead of having a repellent effect. However, recent work showed that DEET acts as an odorant molecule and elicits a behavioural response in the absence of other stimuli. Therefore, DEET must promote some phenomenon connected with the stimuli-sensory system interaction, such as a sensory adaptation, where the sensory system regulates its sensitivity to different stimuli intensities during continuous or repetitive exposure. In this work, we studied different aspects of the insect-DEET interaction through behavioural observations. Previous exposure of fifth instar Rhodnius prolixus nymphs to DEET decreased the behavioural response to this repellent. We observed a decrease in repellence after different times of continuous stimulation with DEET in a time-dependent manner. The response to DEET was recovered 10 min after exposure, when insects were continuously stimulated during 5 or 10 min; maximum repellence was recovered 20 min after exposure when insects were stimulated for 20 min. DEET produced a repellent effect when nymphs were exposed only to its vapours. These results suggest that exposure to DEET produces adaptation in R. prolixus nymphs, and that the behavioural response elicited by DEET occurs via olfaction when no other stimuli are present.

Insecticidal activity of essential oils from eleven Eucalyptus spp. and two hybrids: lethal and sublethal effects of their major components on Blattella germanica.

The aim of the current study was to evaluate the fumigant activity of the essential oils from 11 species of the genus Eucalyptus and two of their hybrids on first instar of Blattella germanica L. The fumigant activity and repellence of the four major monoterpene components of these essential oils also were tested. Fumigant activity was evaluated by exposing nymphs to the vapors emitted by 50 microl of essential oil or monoterpene in a closed container. The lowest knockdown time 50% (KT50) values, expressed in minutes, were elicited by the essential oils of the Eucalyptus grandis X Eucalyptus tereticornis (57.9) hybrid, Eucalyptus sideroxylon A. Cunn (62.0), E. grandis X Eucalyptus camaldulensis (63.8) hybrid, Eucalyptus viminalis Labill (64.1), Eucalyptus dunnii Maiden (64.5), and Eucalyptus grandis (Hill) ex Maiden (68.7). The KT50 values for the remaining essential oils ranged between 74.5 (E. saligna Smith) and 161.4 min (E. tereticornis Smith). The essential oil from the hybrid E. grandis X E. tereticornis was 3.7 times less toxic than dichlorvos (positive control). The KT50 values of monoterpenes were 38.8 for alpha-pinene, 55.3 for 1,8-cineole, 175.6 for p-cymene, and 178.3 for gamma-terpinene. Alpha-pinene was 2.5 times less toxic than dichlorvos. There was a strong positive correlation between the fumigant activity of essential oils and their corresponding 1,8-cineole and alpha-pinene concentration. Repellency was quantified using a video tracking system. Two concentrations of monoterpenes were studied (7 and 70 microg/cm2). All compounds produced a light repellent effect but only when applied at 70 microg/cm2. In all cases, the repellent effect was less than that produced by the broad-spectrum insect repellent N,N-diethyl-3-methylbenzamide (positive control).

Toxicity to vapor exposure and topical application of essential oils and monoterpenes on Musca domestica (Diptera: Muscidae).

The medical and veterinary pest Musca domestica L. has developed resistance to most insecticides used against it. For this reason, there is a constant search for new alternative control tools. The aims of this study were (1) to evaluate the toxicological effects caused by the fumigant activity and the topical application of five essential oils and five monoterpenes in M. domestica adult males and (2) to study the variation of the fumigant activity of the essential oils and monoterpenes according to the solvent used (acetone or a silicone base). Houses flies were exposed to vapors delivered by filter paper treated with 200 microl of essential oil or monoterpene (10%) in acetone or a silicone base. The knockdown time 50% (KT50) values obtained for essential oils (expressed in minutes) were 3.3 (eucalyptus); 10.1 (orange); 10.4 (mint); 10.9 (lavender); and 17.7 (geranium). The KT50 values obtained for monoterpenes (expressed in minutes) were 2.3 (eucalyptol); 7.5 (limonene); 7.6 (linalool); 19.0 (menthone); and 22.6 (menthyl acetate). In all cases, a delay in the onset of poisoning symptoms was observed when a silicone base vehicle was used. When topically applied, the lethal dose 50% (LD50) values for essential oils (expressed in micrograms of oil/insect) were 0.07 (geranium); 0.09 (mint); 0.13 (lavender); 0.14 (eucalyptus); and 0.16 (orange). The LD50 values for monoterpenes (expressed in micrograms of monoterpene/insect) were 0.04 (linalool); 0.09 (menthyl acetate); 0.10 (limonene); 0.11 (menthone); and 0.13 (eucalyptol). These results suggest that the studied essential oils and monoterpenes are potential tools for controlling M. domestica.

Fumigant insecticidal activity and repellent effect of five essential oils and seven monoterpenes on first-instar nymphs of Rhodnius prolixus.

The aim of this study was to evaluate the fumigant and repellent activity of five essential oils (from eucalyptus, geranium, lavender, mint, and orange oil) and seven monoterpenes (eucalyptol, geraniol, limonene, linalool, menthone, linalyl acetate, and menthyl acetate) on first-instar nymphs of the bloodsucking bug Rhodnius prolixus Stahl (vector of Chagas disease in several Latin American countries). Fumigant activity was evaluated by exposing the nymphs to the vapors emitted by 100 microl of essential oil or monoterpene in a closed recipient. The knockdown time 50% (KT50) for eucalyptus essential oil was 215.6 min (seven times less toxic than dichlorvos, a volatile organophosphorus insecticide used as a positive control). The remaining essential oils showed a poor fumigant activity: < 50% of nymphs were knocked down after 540 min of exposure. The KT50 values for monoterpenes, expressed in minutes, were as follows: 117.2 (eucalyptol), 408.7 (linalool), 474.0 (menthone), and 484.2 (limonene). Eucalyptol was 3.5 times less toxic than dichlorvos. No affected nymphs were observed after 540 min of exposure to geraniol, linalyl acetate, or menthyl acetate. Repellency was quantified using a video tracking system. Two concentrations of essential oils or monoterpenes were studied (40 and 400 microg/cm2). Only mint and lavender essential oils produced a light repellent effect at 400 microg/cm2. Geraniol and menthyl acetate produced a repellent effect at both tested concentrations and menthone only elicited an effect at 400 microg/cm2. In all cases, the repellent effect was lesser than that produced by the broad-spectrum insect repellent N,N-diethyl-3-methylbenzamide (DEET).

3-pentanol: a new attractant present in volatile emissions from the ambrosia beetle, Megaplatypus mutatus.

Megaplatypus mutatus (=Platypus mutatus) (Coleoptera: Platypodidae) is an ambrosia beetle that is native to South America. It attacks only standing live trees and causes severe stem breakage and death in commercial poplar (Populus) plantations. Previous work showed that male M. mutatus emits a sex pheromone composed mainly of (+)-sulcatol and sulcatone. We collected male volatile emissions during the hours of maximum emergence by using a specific polar microextraction phase; analyzed the extract by GC-MS; and tested the biological activity of selected compounds in the extract with a walking behavioral assay. Female M. mutatus emerged primarily between 7 and 11 h. In the chemical analyses of volatiles, a third compound, 3-pentanol, was identified in a small percentage of samples. Walking behavioral bioassays with video image analysis showed that at the doses tested, 3-pentanol elicited an attractive response from females.

Decrease in DEET repellency caused by nitric oxide in Rhodnius prolixus.

N,N-diethyl-3-methylbenzamide (DEET) is widely used as an insect repellent; however, little is known about its mode of action. On the other hand, nitric oxide (NO) participates in the olfaction transduction pathway of insects. In this work, nitroso-acetyl-cysteine (SNAC), a nitric oxide donor, or dibutyril-cyclic-GMP (db-cGMP), the cyclic nucleotide analog, were applied on fifth instar nymphs of Rhodnius prolixus before exposing them to DEET, to obtain information about the possible role of NO/cGMP system in the olfaction process. In the first place, we exposed the nymphs to several DEET concentrations (70, 700, 1,750, and 3,500 microg/cm2). All these concentrations produced a repellent effect. A decrease in repellency during the course of the experiment was observed when the nymphs were exposed to high concentrations of DEET (700 and 1,750 microg/cm2), suggesting an adaptation phenomenon. The pre-treatment of the insects with 15 microg /insect of SNAC or 2 microg/insect of db-cGMP produced a reduction of the repellency. An increase in locomotor activity was observed in insects exposed to 350 or 700 microg/cm2 DEET. Although exposure to 70 microg/cm2 DEET produced a high repellency response, it did not modify the insects' locomotor activity. Insects treated with two doses of SNAC before being exposed to 350 microg/cm2 of DEET showed no differences in locomotor activity compared to controls.

Synergism between cis-permethrin and benzoyl phenyl urea insect growth regulators against Aedes aegypti larvae.

Emulsifiable concentrate formulations of 2 insect growth regulator (IGR) benzoyl ureas (novaluron and diflubenzuron) and the pyrethroid cis-permethrin were individually bioassayed against late 3rd- or early 4th-stage larvae of susceptible Aedes aegypti. The emergence inhibition 50% were 0.038, 0.048, and 0.179 ppb for novaluron, diflubenzuron, and cis-permethrin, respectively. We also tested 1:1 (v:v) mixtures of cis-permethrin and benzoyl ureas. Emergence inhibition 50% for novaluron plus cis-permethrin and diflubenzuron plus cis-permethrin were 0.030 and 0.037, respectively, with combination indexes of 0.49 and 0.56, indicating a synergistic interaction between these insecticides. The percentage of larval mortality produced by 0.5 ppb of the mixtures of cis-permethrin and each IGR was compared to the addition of larval mortality produced in separated experiments by 0.25 ppb of each component, after 5, 9 and 13 days of continuous exposure. In all bioassays, the novaluron plus cis-permethrin mixture produced a significantly higher larvicidal effect (P < 0.05) compared with the individual contribution of each compound. The diflubenzuron plus cis-permethrin mixture showed significantly higher larval mortality (P < 0.05) compared with the contributive effect of each insecticide after 9 and 13 days of exposure, but there was no significant difference after 5 days of exposure.