Genetic architecture of the toxicological response to eucalyptol and citronellal in Drosophila melanogaster.

The excessive and indiscriminate use of synthetic insecticides has led to environmental pollution, wildlife destruction, and adverse effects on human health, while simultaneously giving rise to resistance in insect pest populations. This adaptive trait is expressed through various mechanisms, such as changes in the cuticle, heightened activities of detoxifying enzymes, and alterations in the sites of action that reduce their affinity for insecticides. In this context, we associate variation in toxicological response with genomic variation, to identify genetic polymorphisms underlying the different steps of the insect (genotype)-response (phenotype)-insecticide (environment) interaction. Under this framework, our objective was to investigate the genetic factors involved in the toxicological response of D. melanogaster lines when exposed to citronellal and eucalyptol vapors (monoterpenes of plant origin). We quantified KT50 in adult males, representing the time necessary for half of the exposed individuals to be turned upside down (unable to walk or fly). Since the genomes of all lines used are completely sequenced, we perform a Genome Wide Association Study to analyze the genetic underpinnings of the toxicological response. Our investigation enabled the identification of 656 genetic polymorphisms and 316 candidate genes responsible for the overall phenotypic variation. Among these, 162 candidate genes (77.1%) exhibited specificity to citronellal, 45 (21.4%) were specific to eucalyptol, and 3 candidate genes (1.5%) namely CG34345, robo2, and Ac13E, were implicated in the variation for both monoterpenes. These suggest a widespread adaptability in the response to insecticides, encompassing genes influenced by monoterpenes and those orchestrating resistance to the toxicity of these compounds.

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.

Botanical monoterpenes synergise the toxicity of azamethiphos in the vector of Chagas disease, Triatoma infestans (Hemiptera: Reduviidae).

OBJECTIVE: To investigate what toxicological interactions occur when binary combinations of azamethiphos and botanical monoterpenes (eugenol, menthol or menthyl acetate) are applied to Triatoma infestans. METHODS: The toxicity of binary mixtures of azamethiphos and sublethal doses of a monoterpene (eugenol, menthol or menthyl acetate) was evaluated in nymphs of the first stage of T. infestans. Experiments using exposure to filter papers and topical application were carried out. Values of Lethal Concentration 50% (LC50) were calculated in the first case, and values of Lethal Dose 50% (LD50) in the second. RESULTS: The LC50 of azamethiphos applied on filter paper was 50.3 µg/cm2 . However, when it was simultaneously applied with a sublethal concentration of monoterpene, its toxicity increased (LC50 with eugenol = 11.20 µg/cm2 , LC50 with menthyl acetate = 5.30 µg/cm2 , LC50 with menthol = 7.26 µg/cm2 ). When applied topically, the LD50 of azamethiphos was 7.85 µg/insect, but its toxicity drastically increased when it was applied together with sublethal doses of menthol (LD50 = 0.00016 µg/insect) or menthyl acetate (LD50 = 0.00051 µg/insect). The simultaneous application with eugenol did not significantly change azamethiphos toxicity (LD50 = 12.79 µg/insect). CONCLUSIONS: The toxicity of azamethiphos in T. infestans was synergised when it was applied together with eugenol, menthol or menthyl acetate on a filter paper. However, only menthol and menthyl acetate synergysed azamethiphos when mixtures were topically applied. The drastic effects of menthol and menthyl acetate in topical application experiments should be further studied as they could be the basis for developing more efficient triatomicidal products with a lower content of conventional insecticides than those currently used for controlling T. infestans.

OBJECTIF: Etudier les interactions toxicologiques qui se produisent lorsque des combinaisons binaires d'azaméthiphos et de monoterpènes botaniques (eugénol, menthol ou acétate de menthyle) sont appliquées à Triatoma infestans. MÉTHODES: La toxicité de mélanges binaires d'azaméthiphos et de doses sublétales d'un monoterpène (eugénol, menthol ou acétate de menthyle) a été évaluée sur les nymphes du premier stade de T. infestans. Des expériences utilisant une exposition à des papiers filtres et une application topique ont été réalisées. Les valeurs de concentration létale à 50% (CL50) ont été calculées dans le premier cas et les valeurs de dose létale à 50% (DL50) dans le second. RÉSULTATS: La CL50 de l'azaméthiphos appliqué sur papier filtre était de 50,3 µg/cm2 . Cependant, lorsqu'il était appliqué simultanément avec une concentration sublétale de monoterpène, sa toxicité augmentait (CL50 avec eugénol = 11,20 µg/cm2 , CL50 avec acétate de menthyle = 5,30 µg/cm2 , CL50 avec menthol = 7,26 µg/cm2 ). Lorsqu'il était appliqué localement, la DL50 de l'azaméthiphos était de 7,85 µg/insecte, mais sa toxicité augmentait considérablement lorsqu'il était appliqué avec des doses sublétales de menthol (DL50 = 0,00016 µg/insecte) ou d' acétate de menthyle (DL50 = 0,00051 µg/insecte). L'application simultanée d'eugénol n'a pas modifié de manière significative la toxicité de l'azaméthiphos (DL50 = 12,79 µg/insecte). CONCLUSIONS: La toxicité de l'azaméthiphos chez T. infestans a été mise en synergie lorsqu'il a été appliqué avec de l'eugénol, du menthol ou de l' acétate de menthyle sur un papier filtre. Cependant, seuls le menthol et l' acétate de menthyle ont eu un effet synergique avec l'azaméthiphos lorsque les mélanges étaient appliqués localement. Les effets drastiques du menthol et de l' acétate de menthyle dans les expériences d'application topique devraient être plus étudiés car ils pourraient être la base du développement de produits triatomicides plus efficaces avec une teneur inférieure en insecticides conventionnels que ceux actuellement utilisés pour lutter contre T. infestans.

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.

Actividad repelente del aceite esencial de Laurelia sempervirens (Ruiz & Pav.) Tul. (Monimiaceae) en Triatoma infestans (Klug)(Reduviidae) / Repellent activity of the essential oil from Laurelia sempervirens (Ruiz & Pav.) Tul. (Monimiaceae) on Triatoma infestans (Klug) (Reduviidae)

Triatoma infestans (Klug) es el vector principal de la enfermedad de Chagas en Bolivia y los países vecinos. El objetivo de este estudio fue determinar la composición química del AE del laurel chileno, Laurelia sempervirens (Ruiz & Pav.) Tul. (Monimiaceae) y evaluar su efecto repelente en ninfas del quinto estadio de T. infestans. La AE de L. sempervirens se obtuvo por hidrodestilación y se analizó por cromatografía de gases acoplada a espectrometría de masas (CG-EM). Sus componentes principales fueron cis-isosafrol (89.8%), ß-terpineno (3.9%), trans-ocimeno (2.7%) y metileugenol (2.2%). La repelencia se evaluó en un círculo de papel de filtro dividido en dos zonas iguales que se impregnaron con sustancias de prueba [AE o N,N-dietil-3-metilbenzamida (DEET) como control positivo] y acetona como control en blanco, respectivamente. Se analizaron varias concentraciones de sustancias de prueba entre 4.125 y 132 µg/cm2. El AE de L. sempervirens produjo una repelencia significativa a concentraciones iguales o superiores a 66.0 µg/cm2, mientras que DEET repelió a partir de 16.5 µg/cm2. Futuros trabajos serán orientados al estudio de las propiedades repelentes de cis-isosafrol solo y mezclado con ß-terpineno, trans-ocimeno y metileugenol en T. infestans.

Triatoma infestans (Klug) is the principal vector of Chagas disease in Bolivia and neighboring countries. The objective of this study was to determine the chemical composition of the EO of the Chilean laurel, Laurelia sempervirens (Ruiz & Pav.) Tul. (Monimiaceae) and to evaluate its repellent effect on fifth-instar nymphs of T. infestans. The EO from L. sempervirens was obtained by hydrodistillation and analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Their main components were cis-isosafrole (89.8%), ß-terpinene (3.9%), trans-ocimene (2.7%) and methyleugenol (2.2%). Repellency was evaluated on a circle of filter paper divided into two equal zones which were impregnated with test substances [EO or N,N-diethyl-3-methylbenzamide (DEET) as positive control] and acetone as blank control, respectively. Several concentrations of test substances between 4.125 and 132 µg/cm2 were tested. The EO from L. sempervirens produced significant repellency at concentrations equal or above 66.0 µg/cm2, while DEET repelled starting at 16.5 µg/cm2. Future works will be oriented to the study of repellent properties of cis-isosafrole alone and mixed with ß-terpinene, trans-ocimene and methyleugenol on T. infestans.

Eugenol-hyperactivated nymphs of Triatoma infestans become intoxicated faster than non-hyperactivated nymphs when exposed to a permethrin-treated surface.

BACKGROUND: Eugenol is a botanical monoterpene that hyperactivates the blood-sucking bug Triatoma infestans, and permethrin is a pyrethroid with a strong triatomicide effect. In the present work, we tested the hypothesis that eugenol-hyperactivated nymphs of T. infestans pick up more insecticide, and then become intoxicated faster, than non-hyperactivated nymphs when exposed to a permethrin-treated surface. RESULTS: Values of knockdown time 50% (KT50) for third-instar T. infestans exposed to a paper impregnated with permethrin were obtained under the following situations: (a.i.) immediately after topical application of eugenol (KT50: 66.75 min for acetone pre-treated controls, and 46.27 min for eugenol pre-treated nymphs); (a.ii.) 30 min after topical application of eugenol (KT50: 66.79 min for controls, and 66.79 min for eugenol pre-treated nymphs); (b) simultaneously with exposure to eugenol vapors (KT50: 51.90 min for controls, and 39.5 min for nymphs exposed to an eugenol-treated filter paper); and (c) immediately after an injection of eugenol (on average, controls were knocked down after 63.00 min, whereas nymphs injected with eugenol were knocked down after 65.30 min). In other experimental series, the distance traveled (DT) by nymphs exposed to eugenol was quantified in the same situations previously described, but without exposure to permethrin. In (a.i.), the DT in interval 0-30 min after topical application of eugenol was 487.00 (control) and 1127.50 (eugenol) cm; in (a.ii.), the DT in the interval 31-60 min after topical application was 336.75 (control) and 256.75 (eugenol) cm; in (b), DT was 939.08 (control) and 1048.53 (eugenol) cm; and in (c), it was 589.20 (control) and 700.00 (eugenol) cm. The KT50 values for permethrin decreased significantly in situations (a.i.) and (b), and eugenol only produced a significant hyperactivity in the same situations. Finally, the amount of permethrin picked up by non-hyperactivated and hyperactivated nymphs exposed to a film of permethrin was quantified by gas chromatography. Non-hyperactivated nymphs picked up 0.34 µg/insect of permethrin, while hyperactivated nymphs picked up 0.65 µg/insect. CONCLUSION: Results support the hypothesis that eugenol-hyperactivated nymphs of T. infestans pick up more insecticide, and then become intoxicated faster, than non-hyperactivated nymphs when exposed to a permethrin-treated surface.

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.

Effects of chlorpyrifos on enzymatic systems of Cydia pomonella (Lepidoptera: Tortricidae) adults.

The control program of codling moth (Cydia pomonella L.) in the Río Negro and Neuquén Valley is intended to neonate larvae. However, adults may be subjected to sublethal pesticide concentrations generating stress which might enhance both mutation rates and activity of the detoxification system. This study assessed the exposure effects of chlorpyrifos on target enzyme and, both detoxifying and antioxidant systems of surviving adults from both a laboratory susceptible strain (LSS) and a field population (FP). The results showed that the FP was as susceptible to chlorpyrifos as the LSS and, both exhibited a similar chlorpyrifos-inhibitory concentration 50 (IC50 ) of acetylcholinesterase (AChE). The FP displayed higher carboxylesterase (CarE) and 7-ethoxycoumarine O-deethylase (ECOD) activities than LSS. Both LSS and FP showed an increase on CarE activity after the exposure to low-chlorpyrifos concentrations, followed by enzyme inhibition at higher concentrations. There were no significant differences neither in the activities of glutathione S-transferases (GST), catalase (CAT) and superoxide dismutase (SOD) nor in the reduced glutathione (GSH) content between LSS and FP. Moreover, these enzymes were unaffected by chlorpyrifos. In conclusion, control adults from the FP exhibited higher CarE and ECOD activities than control adults from the LSS. AChE and CarE activities were the most affected by chlorpyrifos. Control strategies used for C. pomonella, such as rotations of insecticides with different modes of action, will probably delay the evolution of insecticide resistance in FPs from the study area.

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.

Repellence produced by monoterpenes on Rhodnius prolixus (Hemiptera: Reduviidae) decreases after continuous exposure to these compounds.

Botanical monoterpenes are secondary metabolites present in essential oils produced by plants. Some of them are insect repellents. The bloodsucking bug Rhodnius prolixus Ståhl (Hemiptera: Reduviidae) is one of the main vectors of Chagas disease in the north of South America and some countries in Central America. In this study, we studied the repellence produced by two monoterpenes, menthyl acetate and geraniol, on fifth instar nymphs of R. prolixus. In the absence of other stimuli, both menthyl acetate and geraniol produced a repellent effect from 740 µg/cm(2) and 74 µg/cm(2), respectively. Pre-exposure to each monoterpene reduced the repellent activity produced by the same substance. Additionally, pre-exposure to one monoterpene decreased the behavioral response of the nymphs to the other one. The repellent effect of both monoterpenes also decreased when nymphs' antennae were previously treated with the nitric oxide donor S-nitroso-N-acetyl-cysteine.

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.