| Summary: | Sheep estrous synchronization is mainly based on progestagen-impregnated sponges which could cause vaginitis. Several species of <i>Lactobacillus</i> used as probiotics are commonly used in the treatment or prevention of urogenital infections in humans. However, no studies have been performed to analyze the potential use of probiotics to prevent urogenital infections in sheep. A randomized controlled clinical trial was conducted with 21 one-year-old ewes to develop a model of probiotic infusion in vaginal sponges in order to study their influence in ewe’s vaginal microbiota, general health status, fertility and prolificity. Synchronization of estrus was based on intravaginal sponges for 14 days. Bacterial communities (Enterobacteriaceae and lactic acid bacteria) were highly fluctuating over time and between animals. The safety of probiotic infusion (mix of <i>Lactobacillus</i> spp. 60% <i>L. crispatus</i>, 20% <i>L. brevis</i> and 20% <i>L. gasseri</i>) in the vagina of healthy ewes was firstly confirmed. Neutrophils were observed in 80% (8/10) of the control ewes compared to 36% (4/11) of the ewes in the probiotic group 2 days after sponge removal (<i>p</i> = 0.056). Fertility in the control and probiotic groups was 60% (6/10) and 91% (10/11), respectively <i>p</i> = 0.097. These results suggest that <i>Lactobacillus</i> spp. infusion in the ewe’s vagina does not affect general health status or fertility.
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