| Summary: | Lactobacilli administration has been suggested for the treatment and prevention of bacterial vaginosis, which increases the risk for preterm birth. We aimed to evaluate the vaginal colonization of lactobacilli orally administered to pregnant women at risk for preterm birth. We performed a randomized and controlled crossover study between January 2016 and May 2017. Forty pregnant women at high risk for preterm birth with normal vaginal flora (Nugent score ≤ 3) were randomized to either receive two oral capsules/day each containing 5 × 10<sup>9</sup> <i>Lactobacilli</i> (L.) <i>rhamnosus</i> GR-1 and <i>L. reuteri</i> RC-14 (<i>n</i> = 20) or no treatment (<i>n</i> = 20) for 2 months. Treatments were then crossed over for an additional two months. A vaginal examination and swabbing were performed for assessment of bacterial vaginosis at baseline and every month until study completion. At the same time points, vaginal samples were cultured and subjected to matrix-assisted-laser-desorption/ionization-time-of-flight-mass-spectrometry (MALDI TOF-MS) for the detection of the specific bacterial strains contained in the capsules. The primary endpoint was the presence of the administered lactobacilli strains in the vagina during the first two months of follow-up. Thirty-eight women completed the study. During the first two months of treatment, <i>L. rhamnosus</i> GR-1 was detected in one (5%) woman on the probiotic treatment and 2 (11%) women receiving no treatment (<i>p</i> = 0.6). <i>L. rhamnosus</i> GR-1 was detected in vaginal samples of 4 (11%) women during probiotic treatment (of both groups) and <i>L. reuteri</i> RC-14 was not detected in any samples. The rest of the endpoints were not different between the groups. Altogether, vaginal colonization of lactobacilli following oral administration is low during pregnancy.
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