Promiscuous mating systems are widely distributed among animals and can be promoted by operational sex ratios (number of receptive adults; OSR). In populations where OSR is not biased towards any sex, the possibility that males and females mate with several individuals increases. For both sexes to synchronize in time and space for reproduction, adults should possess or simultaneously acquire nutrients required to reach sexual maturity. Among synovigenic (without a full complement of eggs at eclosion) species, nutrient acquisition, protein in particular, may influence the OSR. In the Agave fly Euxesta bilimeki (Hendel) (Diptera: Ulidiidae), both sexes engage in multiple mating and females frequently expel all or part of the ejaculate. Here, we assessed the effect of protein intake on gonadic development, and estimated OSR from field-collected individuals. Body protein content was compared between wild and laboratory individuals with access to different diets, and mating frequency and individual mating rate were analyzed for cohorts at a 1:1 sex ratio. Both sexes required protein ingestion for gonadic development, but there were no differences in protein content between field-collected males and males fed protein and sugar in the laboratory, despite the fact that males assigned 9.3% of their corporal weight to testicles. Euxesta bilimeki is a promiscuous species where both males and females mate multiply with one or several individuals in short periods of time; thus, large testes size may be linked to the need of voluminous ejaculate production, and might be further exacerbated by female ejaculate expulsion.
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