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The objective of this study was to characterize the vaginal bacterial flora and subsequent pregnancy rates after AI in sheep subjected to 4 different estrous synchronization regimes – the use of two intravaginal devices (silicone or polyurethane sponges), and two different treatment periods (7 or 14 days). Twenty-one multiparous Texel ewes were randomly allocated to 4 treatment groups during the breeding season. In the ewes from Group I (n = 6) and Group II (n = 5), a polyurethane sponge containing 60mgMAP was inserted in the vagina for a period of 7 or 14 days, respectively. In the ewes of Group III (n = 5) and Group IV (n = 5), an intravaginal progesterone releasing insert (IVP4) containing 160mg of progesterone in an inert silicone device, was inserted for 7 or 14 days, respectively. At device withdrawal, each ewe was treated with 200 IU eCG i.m. Standard bacteriological procedures were performed on vaginal mucus samples obtained before the introduction of the devices, at device withdrawal and on the day of AI in all groups. Estrus was recorded with the aid of vasectomized rams every 12 h, and AI was performed 52–54 h after device withdrawal, using fresh semen.