Vaginal secretions are an important source of chemical signals, which affect ewes' attractiveness. Moreover, alterations of vaginal flora reduce sexual attractiveness of estrous ewes. As intravaginal sponges containing progestagens (widely used for estrous synchronization) affect vaginal flora, our aims were to determine if estrous ewes pretreated with intravaginal sponges were less attractive than ewes displaying spontaneous estrus, and if the addition of antibiotic to the sponge mitigated the decreased sexual attractiveness. Seventy-two estrous ewes were used in experiment 1: in 36, estrus was synchronized with commercial intravaginal sponges (50 mg medroxyprogesterone acetate for 14 days, group MAP1), whereas the other 36 were given a PGF2α analogue 19 to 20 days earlier and displayed spontaneous estrus (group C1). In experiment 2, 72 ewes were treated with intravaginal sponges for 14 days; for 36 ewes, the sponges contained 0.02 mg oxytetracycline (group Ox), whereas there was no antibiotic in the sponges for the remaining 36 ewes (group MAP2). In both experiments, sexual attractiveness was determined in 12 groups of six estrous ewes (three MAP1 vs. three C1, and three MAP2 vs. three Ox for Experiments 1 and 2, respectively) located in a 4 × 4 m pen. Courting and mating time that each ram spent with each ewe was recorded. After 5 min, the ewe with which the ram spent more time (most attractive ewe, ranked one, scale one to six) was taken out from the pen. The procedure was repeated until the ram ranked all six ewes, and repeated in the 12 groups in both experiments. In experiment 1, C1 ewes were more attractive than MAP1 ewes (ranks: 2.9 ± 0.3 vs. 4.1 ± 0.3, mean ± SEM, respectively; P < 0.002). In experiment 2, sexual attractiveness of MAP2 and Ox ewes was similar (3.5 ± 0.3 vs. 3.4 ± 0.3, respectively). We concluded that the use of intravaginal sponges impregnated with medroxyprogesterone acetate negatively affected ewes' sexual attractiveness, but this decrease was not mitigated by inclusion of a local antibiotic.