RESUMO
Insecticide resistance is an important factor in the effectiveness of Aedes aegypti control and the related spread of dengue. The objectives of this study were to investigate the status of the organochlorine dichlorodiphenyltrichloroethane (DDT) and pyrethroid (permethrin and deltamethrin) resistance in Trinidad and Tobago populations of Ae. aegypti and the underlying biochemical mechanisms. Nine populations of Ae. aegypti larvae from Trinidad and Tobago were assayed to DDT and PYs using the Centers for Disease Control and Prevention (CDC) time-mortality-based bioassay method. A diagnostic dosage (DD) was established for each insecticide using the CAREC reference susceptible Ae. aegypti strain and a resistance threshold (RT), time in which 98-100% mortality was observed in the CAREC strain, was calculated for each insecticide. Mosquitoes which survived the DD and RT were considered as resistant, and the resistance status of each population was categorised based on the WHO criteria with mortality <80% indicative of resistance. Biochemical assays were conducted to determine the activities of α and ß esterases, mixed function oxidases (MFO) and glutathione-S-transferases (GST) enzymes which are involved in resistance of mosquitoes to DDT and PYs. Enzymatic activity levels in each population were compared with those obtained for the CAREC susceptible strain, and significant differences were determined by Kruskal-Wallis and Tukey's non-parametric tests (P<0.05). The established DDs were 0.01 mg l(-1), 0.2 mg l(-1) and 1.0 mg l(-1) for deltamethrin, permethrin and DDT, respectively; and the RTs for deltamethrin, permethrin and DDT were 30, 75 and 120 min, respectively. All Ae. aegypti populations were resistant to DDT (<80% mortality); two strains were incipiently resistant to deltamethrin and three to permethrin (80-98% mortality). Biochemical assays revealed elevated levels of α-esterase and MFO enzymes in all Ae. aegypti populations. All, except three populations, showed increased levels of ß-esterases; and all populations, except Curepe, demonstrated elevated GST levels.Metabolic detoxification of enzymes is correlated with the manifestation of DDT and PY resistance in Trinidad and Tobago populations of Ae. aegypti. The presence of this resistance also suggests that knock down (kdr)-type resistance may be involved, hence the need for further investigations. This information can contribute to the development of an insecticide resistance surveillance programme and improvement of resistance management strategies aimed at combatting the spread of dengue in Trinidad and Tobago.
Assuntos
Aedes , DDT , Inseticidas , Piretrinas , Animais , Resistência a Inseticidas , Larva , Trinidad e TobagoRESUMO
Anopheles pseudopunctipennis, one of the primary vectors of malaria in the southwest of Colombia, was evaluated for susceptibility to the 3 major insecticide groups (organophosphates, pyrethroids, and carbamates) by bioassay and biochemical assay. Larval populations, which were collected principally from irrigation channels in agricultural areas, where the intensity of insecticide use varied, were utilized to establish susceptibility for the 1st time in this species. The baselines for each population showed a range of biological susceptibility to the insecticides evaluated, but overall no resistance was detected according to standards established by the World Health Organization. The high sensitivity of biochemical microassays enabled the detection of a small proportion of mosquitoes with higher levels of nonspecific esterases and mixed-function oxidases from 2 areas where agricultural application of organophosphate and pyrethroid insecticides had been heavy. These differences were not sufficient to affect susceptibility as measured by bioassay. No evidence of insensitive acetylcholinesterase was observed. Absence of resistance in areas that have experienced heavy insecticide application could be explained by genetic drift, by gene flow from areas without insecticide pressure, by manner of exposure to the insecticides, or by recent changes in agricultural activities that decreased insecticide use. Baseline values were established that serve as provisional susceptibility thresholds for applying simple Centers for Disease Control and Prevention biochemical assay and bioassay methods to larvae of this anopheline species.
Assuntos
Anopheles , Inseticidas , Controle de Mosquitos , Acetilcolinesterase/análise , Animais , Anopheles/enzimologia , Bioensaio , Colômbia , Esterases/análise , Malation , Controle de Mosquitos/métodos , Oxirredutases/análise , Permetrina , Propoxur , PiretrinasRESUMO
Isofemale lines were established containing either, both, or neither of the elevated esterase and oxidase resistance mechanisms conferring pyrethroid resistance in a Guatemalan strain of Anopheles albimanus (Wiedemann). Plots of esterase and oxidase levels for individual mosquitoes from these single families correlated with data obtained using oxidase and esterase synergists in bioassays run in the bottle format. Mixed populations of pyrethroid-resistant A. albimanus adult females were selected using DDT, permethrin, or malathion; and the esterase and oxidase levels of the individual progeny were plotted. These data showed that the 3 classes of insecticide selected the 2 mechanisms differently. These results are discussed in terms of the problem of multiresistance surveillance in the field, especially concerning pyrethroid insecticides and the interaction of agricultural and public health insecticide application.
Assuntos
Anopheles , Controle de Mosquitos , Piretrinas , Animais , Anopheles/enzimologia , DDT , Esterases/metabolismo , Feminino , Guatemala , Resistência a Inseticidas , Malation , Controle de Mosquitos/métodos , Oxirredutases/metabolismo , PermetrinaRESUMO
Reduced fenitrothion susceptibility in Haitian Anopheles albimanus is documented using time/mortality measurements from otherwise standard World Health Organization (WHO) bioassays. Survival beyond a time threshold in bioassays is shown to be highly correlated with elevated non-specific esterase levels. A shift in resistance incidence from less than 20 to over 60% in a six-month period is documented using both the bioassay and microassay procedures, showing the potential of microplate assay methods in early detection of resistance. Conventional 24-hour reading of WHO bioassay data failed to detect resistance until the level reached 60%. Resistance appeared to be focal, with an increase in intensity coinciding with a fenitrothion spray cycle for malaria control.
Assuntos
Anopheles , Fenitrotion , Animais , Bioensaio , Resistência a Medicamentos , HaitiRESUMO
Simple microplate assay methods for determining the frequency of insecticide resistance in single mosquitos were used to study the distribution and localization of organophosphate and carbamate resistance in field populations of Anopheles albimanus Weidemann in Guatemala, where such resistance, caused by heavy use of agricultural pesticides, has long been assumed to be widespread. Areas of complete susceptibility to organophosphates and carbamates were observed, as well as areas where the resistant phenotypes represented up to 98% of the population. Overall, the resistance levels were lower and more localized than expected. Two mechanisms of resistance were identified by the microassay methods. These were the elevated esterase (nonspecific esterase) and insensitive acetylcholinesterase mechanisms which were selected independently, the former (documented for the first time in Central American anophelines) being predominant. These methods represent a promising new technology for the detection and assessment of resistance and will facilitate improved control strategy decisions.
Assuntos
Anopheles/efeitos dos fármacos , Carbamatos , Inseticidas , Compostos Organofosforados , Acetilcolinesterase/análise , Animais , Anopheles/enzimologia , Esterases/análise , Guatemala , Resistência a Inseticidas/genética , FenótipoRESUMO
A straightforward method is described for determination of the activities of erythrocyte acetylcholinesterase and plasma cholinesterase from single 10-mul samples of highly diluted whole blood. The procedure is a spectrophotometric microassay adapted for use with microtitration plates. The use of an enzyme immunoassay reader facilitates the rapid analysis of large numbers of samples. An inexpensive adaptation of the method was also devised that requires no instrumentation and was evaluated under field conditions in Haiti. Either the field or laboratory versions of the method can accurately determine increments of 10% or less in the level of inhibition of acetylcholinesterase or cholinesterase in whole blood, plasma, or erythrocytes.
Assuntos
Colinesterases/sangue , Eritrócitos/enzimologia , Acetilcolinesterase/sangue , Agricultura , Haiti , Humanos , Microquímica/métodos , Espectrofotometria/métodosRESUMO
Simple microplate assay methods for determining the frequency of insecticide resistance in single mosquitos were used to study the distribution and localization of organophosphate and carbamate resistance in field populations of Anopheles albimanus Weidemann in Guatemala, where such resistance, caused by heavy use of agricultural pesticides, has long been assumed to be widespread. Areas of complete susceptibility to organophophates and carbamates were observed, as well as areas where the resistant phenotypes represented up to 98 percent of the population. Overall, the resistance levels were lower and more localized than expected. Two mechanisms of resistance were identified by the microassay methods. These were the elevated esterase (nonspecific esterase) and insensitive acetylcholinesterase mechanisms which were selected independently, the former (documented for the first time in Central American anophelines) being predominant. These methods represent a promising new technology for the detection and assessment of resistance and will facilitate improved control strategy decisions