The sexual compatibility between laboratory (LF) and wild (WF) strains of the Mexican fruit fly, Anastrepha ludens (Loew), was analyzed using analogous methodologies and experimental arenas under both laboratory and field conditions. Sexual compatibility was quantified with the following indices: the isolation index (ISI), male relative performance index (MRP), female relative performance index (FRPI), and the relative sterility index (RSI). ISI detected a certain level of incompatibility between strains under both laboratory and field conditions, because LF females tended to mate with LF males. LF mating performance was higher under laboratory than under field conditions. The relative performance indices for LF and the relative sterility index were higher in the laboratory than in the field. Differences between LF and WF in the times that males started calling and mating were observed in both environments. Importantly, WF males reduced their sexual activity under laboratory environments, whereas LF maintained similar activity levels in both conditions. The possible applications of the above-mentioned methods, not only to assess fly quality but also to determine the suitability of conditions in mass-rearing facilities, are discussed. Correlating laboratory quality to sexual behavior may contribute in the development of environmental parameters for mass-rearing facilities.