Maternal gut and breast milk (BM) are key in vertically transmission bacteria to infants, shaping their gut microbiota in early life. Although the establishment of early gut microbiota is known, the role of the combined influence of maternal factors and newborn characteristics is not explored. In this study, we aimed to assess the influence of maternal BMI and total body fat, age, delivery mode, and newborn sex on the diversity and composition of the BM and gut microbiota (GM) in mother–newborn dyads. In this cross-sectional study, of the 986 pregnant women candidates, 53 participated, and, finally, 40 mother–newborn dyads exclusively breastfeeding at 20–28 days postpartum were included. Metataxonomic profiling of DNA extracted from BM and fecal samples was conducted using 16S rRNA sequencing. Globally, the findings offer valuable insights that excessive adiposity, age, and C-section delivery influence a lower abundance of specific taxa in the BM, maternal gut, and gut of newborns. Also, the simultaneous analysis of maternal factors and newborn characteristics shows that maternal age and newborn sex explain an important variation in the microbiota composition. These results add to the understanding of the intricate interplay between maternal factors and the microbial communities that influence early-life gut and BM microbiota.