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.

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.

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.

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.

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

N,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 f ifth-instar nymphs of the blood-sucking bug Rhodnius prolixus Sta˚l (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, 74mg/cm2. Nymph age (between 1­3h and 99d from last ecdysis) had no influence on repellency. Topical application of 750mg of DEET per nymph produced a mortality rate between 0% (24h after application) and 40% (7d 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.