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Background: Infantile colic has a multifactorial etiology; although various treatments have been attempted to manage and alleviate its symptoms, a solution is lacking, adversely affecting mothers and their babies. Recent studies have suggested that dysbiosis may play a role in the pathogenesis of infantile colic and that modulating the gut microbiota, including the use of probiotics, may aid its management.Purpose: This single-blind randomized controlled study evaluated the effect of probiotics (Actiregularis, 5 × 10 6 ) added to the diet of mothers on infantile colic symptoms and neonatal gut microbiota content.Methods: A probiotic drink containing the Actiregularis (5 × 10 6 ) strain was added to the diet of mothers in the experimental group once daily for 15 consecutive days. Stool samples were collected from each infant twice, on days 0 and 15, and fecal 16s rRNA gene sequencing and compositional-based metabolomic analyses were performed. The mothers recorded the babies' crying frequency and severity for 15 days using a daily form created by the researchers. This study was registered at ClinicalTrials.gov (ID: NCT04374955).Results: Infants whose mothers were supplemented with Actiregularis for 15 days showed a decreased frequency (p=0.00) and intensity (p<0.001) of crying as well as a significantly increased bacterial diversity in the stools (p=0.017). This variety was substantially affected by the addition of probiotic products. The greatest species diversity was observed in the group treated with probiotics, while the least diversity was observed in the control group (Shannon, p=0.0043; Simpson, p=0.017). Conclusion:Babies treated with Actiregularis added to their mother's diet for 15 days showed decreased crying frequency and intensity and increased bacterial diversity and density.

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