Fluorescent dyes are commonly used in the sterile insect technique (SIT) for marking insects for a proper identification after recapture. However, the quality of the mark must be balanced against insect performance, because dyes can negatively affect some parameters of insect performance and reduce their effectiveness in control with the SIT. We determined the visibility and persistence and the effect of dyes on the quality of Anastrepha obliqua (Macquart) and Anastrepha ludens (Loew) (bisexual and genetic sexing strains) by testing four concentrations of a dye (Day-Glo) from 0 to 2.5 g dye/kg of pupae. Visibility and persistence of the mark were positively affected by dose and negatively affected by the length of time the samples were kept in a solution of 75% alcohol. However, upon dissection, even the lowest dose of dye was visible under a fluorescence microscope. Between dyed and undyed pupae (control), no significant differences were observed in rates of emergence, fliers and flight ability, and survival in two tests, with water and without food and without water and food, at any of the concentrations tested. Furthermore, no significant difference in mating competitiveness was detected between control pupae and those dyed at 1.0 and 2.5 g dye/kg pupae. We discuss our results with the possibility of reducing the dose of dye in these three flies, because the heads are large enough to capture sufficient particles to permit identification with the current methods of detection.