Experimental evolution is a powerful approach to study the mechanisms underlying the adaptation of selected characters under the conditions chosen in the laboratory. Drosophila melanogaster is a species frequently used to investigate the experimental evolution of characters, especially those related to reproduction. Recent intra-generational studies showed that cis-vaccenyl acetate (cVa), a sex pheromone transferred with bacteria on eggs by females either 1 day (D1) or 5 days (D5) after copulation, differentially affected the behavior and pheromone release in adult males emerging from these eggs. Here, we extended this finding to determine whether this alternative egg exposure repeated over many generations could affect a larger set of reproduction-related characters in both sexes. To test the repetitive effects of maternal D1 or D5 post-copulatory factors, we carried out an experimental selection procedure consisting of exposing eggs during 40 successive generations to D1 or D5 maternal post-copulatory factors. We compared cVa and cuticular pheromones, courtship and mating behaviors, and fecundity at different generations in flies of D1 and D5 lines. Based on findings obtained at earlier generations, we also determined survival, bacterial composition and gene expression in adults. Some of these complex traits significantly diverged between D1 and D5 lines indicating that maternal post-copulatory factors transmitted to eggs can influence adult life history traits.