A simple and affordable method for estimating the fluid volume a mosquito sucks using food dyes

Abstract

Background: Blood-sucking by mosquitoes is an inevitable behavior when pathogens are transmitted among humans. Adenine nucleotides such as ATP are known as phagostimulants for mosquitoes and are widely used to induce and enhance the blood-sucking activity in an artificial manner. Although using ATP solution is convenient to introduce a variety of substances (for example chemicals and pathogens) into the mosquito body via sucking, establishing an easy and cost-effective method to quantify the amount of solution ingested has yet to be reported. Results: A set of commercial food dyes (green, blue, yellow, and red) was employed in this study. Each dye was added to ATP solution used to colorize the abdomen of Ae. aegypti female mosquitoes after ingestion. The intake of food dyes did not show any toxicity to the mosquitoes, affecting neither ATP-sucking behavior nor survival of the mosquitoes. We observed that quantifying the color intensity of green dye in the mosquito abdomen by spectral analysis, as well as distinguishing the size of the colored abdomen using the naked eye, allowed the estimation of the amount of ingested solution. Using this method, capsaicin, a pungent component of chili peppers, was identified as an aversive tastant that can discourage mosquitoes from sucking the ATP solution. Conclusions: Employing commercially available, non-toxic food dyes converted ATP-driven membrane feeding into an easy-to-use method to estimate the amount of solution ingested by mosquitoes. This method can be further applied for a variety of experiments such as introducing a certain quantity of chemical compounds or microbes into the mosquito body.