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The  Drosophila  model has become a leading platform for investigating mechanisms that drive feeding behavior and the effect of diet on physiological outputs. Several methods for tracking feeding behavior in flies have been developed. One method, consumption-excretion or Con-Ex, provides flies with media labeled with dye and then quantifies the amount of dye excreted into the vial as a measure of consumption. We previously found that Blue 1 and Orange 4 work well in Con-Ex and can be used as a dye pair in food preference studies. We have expanded our development of Con-Ex by identifying two additional dyes, Orange G and Yellow 10, that detect the anticipated effects of mating status, strain, starvation and nutrient concentration. Additionally, Orange G and Yellow 10 accumulate linearly in excretion products out to 48 h and the excreted volumes of these two dyes reflect the volumes consumed. Orange G also works with Blue 1 as a dye pair in food preference studies. Finally, consumption of Blue 1, Orange 4, Orange G or Yellow 10 does not affect ethanol sedation or rapid tolerance to ethanol. Our findings establish that Orange G and Yellow 10, like Blue 1 and Orange 4, are suitable for use in Con-Ex.

Identification of additional dye tracers for measuring solid food intake and food preference via consumption-excretion in Drosophila