Increased activity of cytochrome P450 monooxygenase enzymes in reference insecticide-resistant Aedes aegypti strains from Cuba

Leidys French Pacheco; María Magdalena Rodríguez Coto; Juan Andrés Bisset Lazcano; Yanelys Ricardo Leyva; Gladys Gutiérrez Bugallo; Ilario Fuentes López

2013, Number 3

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Rev Cubana Med Trop 2013; 65 (3)

**Increased activity of cytochrome P450 monooxygenase enzymes in reference insecticide-resistant Aedes aegypti strains from Cuba**

French PL, Rodríguez CMM, Bisset LJA, Ricardo LY, Gutiérrez BG, Fuentes LI

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Language: Spanish
References: 23
Page: 328-338
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ABSTRACT

Introduction: cytochrome P450 monooxygenase detoxifying enzymes (MFO) are one of the main resistance mechanisms of Aedes aegypti to insecticides. In vivo studies of the presence of these enzymes have been conducted in Cuba with the use of synergists. However, their activity has not been quantitatively determined in vitro, an indispensable step in studies about metabolic resistance in insects.
Objective: standardize a method to detect the activity of cytochrome P450 monooxygenase in vitro, and then determine such activity in larvae and adults of Aedes aegypti reference strains.
Methods: the study was based on three laboratory strains of Aedes aegypti selected for 14 or 15 generations with temephos, deltamethrin or propoxur, respectively, and a strain susceptible to insecticides.
Results: the conditions for enzyme activity assays were established (protein and substrate concentration: 0.4 mg/mL and 12 mmol/L, respectively, and reaction time: 10 min). There was a significant increase in cytochrome P450 monooxygenase activity in resistant strains, with a higher phenotypic frequency in the larval stage.
Conclusions: modifications to the technique used for determination of enzymatic activity made it possible to distinguish between mosquitoes from susceptible and resistant strains in larval and adult stages, providing a new tool for the detection of metabolic resistance in Cuba.

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