Recently there has been considerable debate in the scientific community regarding the placenta as the host of a unique microbiome. No studies have addressed the associations of clinical conditions such as maternal obesity, or localizations on the placental microbiome. We examined the placental microbiome in a multi-ethnic maternal pre-pregnant obesity cohort using controls for environmental contaminants and an optimized microbiome protocol to enrich low bacterial biomass samples. We confirmed that a distinct placenta microbiome does exist, as compared to the environmental background. The placenta microbiome consists predominantly of Lactobacillus, Enterococcus and Chryseobacterium. Moreover, the microbiome in the placentas of obese pre-pregnant mothers are less diverse when compared to those of mothers of normal pre-pregnancy weight. Lastly, microbiome richness also decreases from the maternal side to fetal side. In summary, our study reveals associations of placental microbiome with placenta biogeography and with maternal pre-pregnant obesity.