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.

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.

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.

High effectiveness of an adulticide-larvicide formulation for field control of sandflies (Diptera: Psychodidae) in the city of Clorinda, Argentina.

In Argentina, Leishmania infantum (syn. L. chagasi) is the etiologic agent of human visceral leishmaniosis (HVL), and Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) is the main vector. The objective of this study was to evaluate the effectiveness and residual effect of two commercial insecticide formulations, one with permethrin and pyriproxyfen as active ingredients (Dragon Max®) and the other with only permethrin (Flop®) for the control of sandflies. Both formulations were applied in chicken coops and other surroundings structures of the peridomicile of urban houses in Clorinda, Formosa (Argentina). Entomological monitoring was carried out weekly for 44 weeks after the intervention. The results showed great effectiveness and residual effect up to 21 weeks post-intervention for Dragon Max®. This result could be explained by the excellent larvicidal activity of the Insect Growth Regulator (IGR) pyriproxyfen against the immature forms of phlebotomines and by the delay on the restoration of the natural threshold of the vector population in treated sites.

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.

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.

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.

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.

Aedes aegypti (Diptera: Culicidae): evaluation of natural long-lasting materials containing pyriproxyfen to improve control strategies.

Natural materials such as beeswax or a paraffin/stearin mixture containing pyriproxyfen and used as a slow release formulation may convert any breeding place into a larvicidal ovitrap for Aedes aegypti (L.) control. Effectiveness and residual activity of beeswax and paraffin/stearin 1:1 discs containing from 10(-5) to 10(-1) % pyriproxyfen and sticked at the bottom of plastic jars were evaluated for adult emergence inhibition (EI) on late 3rd or early 4th instar A. aegypti larvae. At the initial time t = 0, the EI was 100% for vessels containing beeswax or paraffin/stearin 1:1 discs containing up to 10(-4)% pyriproxyfen. For the lowest pyriproxyfen concentration of 10(-5)%, paraffin/stearin mixture gave a higher EI% value than beeswax (100 and 50%, respectively). Jars were kept at room temperature, and water was totally replaced every 15 days. Bioassays for residual activity repeated monthly showed that at 30 days and for pyriproxyfen 10(-5) % and both matrices, the EI values were low and comparable to control values. For pyriproxyfen 10(-4) %, EI remained above 95% for at least 90 days and around 75% up to 180 days. The EI values are always higher for paraffin/stearin mixture than for beeswax. For all other higher concentrations, 100% EI was obtained at least during 300 days. In a semi-field trial, paraffin/stearin/sand O-rings (2:1:2), containing pyriproxyfen 1%, were sunken in 200-l water-storage tanks and held outdoors in a shadow place. After 72 h, a 250-ml aliquot was taken (t = 0) obtaining 100 % EI. Water level was completed to 200 l every 15 days and bioassays repeated monthly as before. Residual activity remains with 100% EI at least for 6 months.

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.

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.

Knockdown and larvicidal activity of six monoterpenes against Aedes aegypti (Diptera: Culicidae) and their structure-activity relationships.

The relationships between physicochemical parameters of majority components of Eucalyptus essential oils and their insecticide effect were evaluated on Aedes aegypti (L.) (Diptera: Culicidae). The octanol-water partition coefficients of the monoterpenes were estimated by the atom/fragment contribution method and the vapor pressures were determined by our laboratory in previous studies. The larvicidal activity (LC50 (ppm)) and knockdown effect (KT50 (min)) of each component was determined. The results show that the toxicity of EOs main components of Eucalyptus on adults and larvae of A. aegypti is strongly related to their physicochemical properties (vapor pressure and Log P). However, the interaction of both variables (vapor pressure * Log P) explains the toxicological phenomenon more precisely. The regression models were expressed as follows: KT 50(min) = - 10.9 + 3.7 * Log P + 1.9 * 1/Pvapor (R(2) = 0.80; F = 42.5) and LC 50(ppm) = - 94.3 + 438.6 * 1/Log P + 2.8 * 1/Pvapor (F = 57.8; R(2) = 0.85). The six evaluated components present different functional groups. Therefore, it was considered to evaluate the monoterpenes as a group and separated in two groups: oxygenated monoterpenes (α-terpineol, 4-terpineol, and 1,8-cineole) and terpene hydrocarbons (γ-terpinene, p-cymene, and α-pinene). The results show the regression models for each group as follows: (A) oxygenated terpenes: KT 50(min) = - 515.3 + 1613.2 * 1/Log P + 5, 2 * 1/Pvapor (F = 3176.7 R(2) = 0.99) and LC 50(ppm) = - 1679.4 + 5402.1 * 1/Log P + 12.7 * 1/Pvapor (F = 282.9; R(2) = 0.99). (B) Hydrocarbons terpenes: KT 50(min) = 18.2 - 58.3 * 1/Log P + 2.7 * 1/Pvapor (F = 171.7; R(2) = 0.97) and LC 50(ppm) = - 21.1 + 174.9 * 1/Log P - 14.3 * 1/Pvapor (F = 410.0; R(2) = 0.99). The association between the toxic effect of the evaluated monoterpenes against A. aegypti and the physicochemical properties can be better described when they are separated into functional groups (hydrocarbons vs. oxygenated terpenes).

A new ovitrap made of slow release natural materials containing pyriproxyfen for Aedes aegypti (Diptera:Culicidae) control.

ABSTRACT This initial study is aimed to measure the performance of incorporating pyriproxyfen in natural materials with low environmental impact to obtain slow release formulations that can be used as larvicidal or autocidal ovitraps avoiding hatched Aedes aegypti (L.) eggs to emerge as adults. Hollow candles made of beeswax or paraffin:stearin 1:1 mixture containing pyriproxyfen 0.01 and 0.05% were prepared and used as holding water containers for larval bioassay. Pyriproxyfen was released quickly into the larvae-breeding water. Ae. aegypti larvae were introduced immediately after the addition of tap water to the hollow candles (t = 1 min) or after 1, 4, and 8 h. More than 40% of the larvae did not emerge as adults for t = 1 min, reaching 80-100% when the larvae were added after 1 or 4 h, respectively. The hollow candles were kept at room temperature, and water was replaced every 15 d. Bioassays performed every 30 d showed that the residual activity obtained for both matrices and both concentrations of pyriproxyfen was higher than 360 d, with 100% inhibition of adult emergence.

Effect of release rate and enantiomeric composition on response to pheromones of Megaplatypus mutatus (Chapuis) in poplar plantations of Argentina and Italy.

Megaplatypus mutatus (=Platypus sulcatus Chapuis) is an Ambrosia beetle native to South America, which was recently introduced in Italy and its presence there is causing severe damage to the local poplar plantations. The male M. mutatus pheromone is composed of (S)-(+)-6-methyl-5-hepten-2-ol [(+)-sulcatol], 6-methyl-5-hepten-2-one (sulcatone) and 3-pentanol. A series of field trials testing dose, blend and enantiomer composition performed in Argentina and Italy evaluated attraction and found that the optimal release rate of pheromone components as baits in cross vane baited traps (CIPEIN-CV) was 6, 6 and 30 mg day−1 of sulcatone, (+)-sulcatol and 3-pentanol, respectively. It was also determined that racemic sulcatol is as effective as the pure (+)-isomer for the purpose of beetle catch, due to the inert nature of the (−)-isomer allowing the usage of low cost racemic sulcatol instead of highly expensive (+)-sulcatol. The results of our work contribute to the development of pheromone-based local technologies with low environmental impact and low cost for control or monitoring of an important pest.

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.

Post-control surveillance of Triatoma infestans and Triatoma sordida with chemically-baited sticky traps.

BACKGROUND: Chagas disease prevention critically depends on keeping houses free of triatomine vectors. Insecticide spraying is very effective, but re-infestation of treated dwellings is commonplace. Early detection-elimination of re-infestation foci is key to long-term control; however, all available vector-detection methods have low sensitivity. Chemically-baited traps are widely used in vector and pest control-surveillance systems; here, we test this approach for Triatoma spp. detection under field conditions in the Gran Chaco. METHODOLOGY/PRINCIPAL FINDINGS: Using a repeated-sampling approach and logistic models that explicitly take detection failures into account, we simultaneously estimate vector occurrence and detection probabilities. We then model detection probabilities (conditioned on vector occurrence) as a function of trapping system to measure the effect of chemical baits. We find a positive effect of baits after three (odds ratio [OR] 5.10; 95% confidence interval [CI(95)] 2.59-10.04) and six months (OR 2.20, CI(95) 1.04-4.65). Detection probabilities are estimated at p ≈ 0.40-0.50 for baited and at just p ≈ 0.15 for control traps. Bait effect is very strong on T. infestans (three-month assessment: OR 12.30, CI(95) 4.44-34.10; p ≈ 0.64), whereas T. sordida is captured with similar frequency in baited and unbaited traps. CONCLUSIONS/SIGNIFICANCE: Chemically-baited traps hold promise for T. infestans surveillance; the sensitivity of the system at detecting small re-infestation foci rises from 12.5% to 63.6% when traps are baited with semiochemicals. Accounting for imperfect detection, infestation is estimated at 26% (CI(95) 16-40) after three and 20% (CI(95) 11-34) after six months. In the same assessments, traps detected infestation in 14% and 8.5% of dwellings, whereas timed manual searches (the standard approach) did so in just 1.4% of dwellings only in the first survey. Since infestation rates are the main indicator used for decision-making in control programs, the approach we present may help improve T. infestans surveillance and control program management.

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.