Activity of increased specific and non-specific esterases and glutathione transferases associated with resistance to permethrin in pediculus humanus capitis (phthiraptera: pediculidae) from Argentina.

Enhanced metabolism by oxidative enzymes is a major cause of pyrethroid resistance in insects. In this work, we evaluated the role of specific and non-specific esterases in head louse populations from Buenos Aires with different levels of resistance to permethrin. As esterase activity is substrate-dependent, four different esters were used as unspecific substrates in order to obtain a better characterization of the possible role of these enzymes in the resistance phenomenon. The unspecific substrates were phenylthioacetate, 1- and 2-naphtyl-acetate, and p-nitrophenyl acetate. A 7-coumaryl permethrate was synthesized and used as a specific substrate to measure pyrethroid esterases by a very sensitive microfluorometric method. The results on pyrethroid esterase activity obtained with this substrate showed that these enzymes contribute to the detoxifying activity in resistant populations, although no correlation was found between pyrethroid esterase activity and resistance ratios. In this study, we established that the activity of esterase against specific and non-specific substrates is increased in pyrethroid-resistant populations of head lice from Buenos Aires. Also, dichlorodiphenyltrichloroethane (DDT) resistance values demonstrated that there is a DDT cross-resistance phenomenon in pyrethroid-resistant head louse populations and suggested that an alteration in the receptor of the nervous system (kdr gen) is a key factor of the resistance phenomena in these head louse populations.

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

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

7-Coumaryl permethrate and its cis- and trans-isomers as new fluorescent substrates for examining pyrethroid- cleaving enzymes.

New esterase substrates were synthesized using cis-, trans- and cis-trans-permethrinic acid chloride and then used to measure pyrethroid-cleaving enzymes in insects. The new substrates, namely cis-, trans- and cis-trans-7-coumaryl permethrates (7-CP), show a structure very similar to permethrin insecticide and yield fluorescent products on hydrolysis. These substrates were hydrolyzed by a commercial porcine preparation that provided esterase-specific activity, and were stable at different pH values (5.2-7.8). Studies made with house fly, Musca domestica (L.), homogenates showed that these compounds are appropriate for determining pyrethroid hydrolysis activity on individual insects. The measured activity of house fly esterase was 870 relative fluorescence units (RFU) min(-1) with cis-7-CP as substrate, 1117 RFU min(-1) with trans-7-CP as substrate and 1423 RFU min(-1) with cis-trans-7-CP as substrate. The fluorescent substrates for pyrethroid-cleaving esterases described in this paper have advantages over methods already given in the literature. They are substrates with structures very similar to pyrethroids, the cleavage of which can be followed by an increase in fluorescence emission at 440 nm; it takes only about 5 min to measure the reaction, and moreover the high sensitivity of the fluorescence technique allows the quantification of esterase activity on individual insects.

Thermal behaviour, biological activity and conformational study of a [methoprene/beta-cyclodextrin] complex in a smoke generating formulation.

Methoprene, an insect growth regulator, was complexed with beta-cyclodextrin, yielding a stable inclusion complex. TGA, X-ray powder diffraction and conformational analysis have been used to confirm the nature of this inclusion complex. The interaction between methoprene and beta-cyclodextrin was investigated by means of Molecular Mechanics. The results account for the formation of a 1:1 inclusion complex stabilised by Van der Waals forces and hydrogen bonds. The [methoprene-beta-cyclodextrin] complex included in smoke generating formulations and protected from thermal decomposition by the foaming agent azodicarbonamide was shown to be stable enough to release methoprene in fumes with good yields. The improved stabilty of the methoprene complex showed a correlation with increased biological activity against Musca domestica.

Attractant volatiles released by female and male Triatoma infestans (Hemiptera: Reduviidae), a vector of chagas disease: chemical analysis and behavioral bioassay.

Volatiles emitted by male and female T infestans before and during copula were collected on Porapak-Q filters, desorbed with dichloromethane, and analyzed by gas chromotography and gas chromatography-mass spectrometry after confirmation of attractiveness in an arena bioassay. Chemical analysis confirmed the presence of (R,S) -2- and 3-methylbutan-1-ol in a 2:1 ratio; short chain acids (ethanoic to nonanoic acid); long chains acids decanoic to (Z)-9-octadecenoic acid; aliphatic aldehydes (hexanal to nonanal), benzaldehyde and dipropylsulphide from insects in copula. Electroantennographic studies conducted with a homologous series of aliphatic aldehydes on female and male T infestans showed that, for a given dose, EAG responses elicited from both sexes increased with increased chain length up to nonanal, after which EAG-activity declined. Attractiveness of non-acidic trace components identified in the volatiles were tested on male and female T. infestans, in an arena bioassay using a video tracking method. Aliphatic C6 to C10 aldehydes were tested: hexanal (1-100 microg) and heptanal (10 microg) were attractive to female T. infestans, high doses of octanal and nonanal (1-100 microg) were Unattractive to male and female T. infestans but low doses of nonanal (0.01-0.1 microg) were attractive to male T infestans. Benzaldehyde was highly attractive to female T. infestans at low doses (0.05- 0.1 microg). 3-methylbutan-1-ol was attractive to male T infestans at high dose (1,000 microg). (S) or (S,R) 2-methyl-butan-1-ol were attractive to males or females (1-1,000 microg). Blends of hexanal and benzaldehyde (20:1 and 40:1) showed an additive effect on attraction compared with hexanal alone, when tested on female T. infestans. The study has demonstrated the presence of a number of electrophysiologically and behaviorally active compounds in volatiles emitted by T. infestans in copula that may have a role in the postulated copulation pheromone.