Examining the relationship between maternal body size, gestational glucose tolerance status, mode of delivery and ethnicity on mother’s milk microbiota at three months post-partum

Background: Few studies have examined how maternal body mass index (BMI), mode of delivery and ethnicity affect the microbial composition of human milk and none have examined associations with maternal metabolic status. Given the high prevalence of maternal adiposity and impaired glucose metabolism, and the importance of human milk in the colonization of the infant gut, we systematically investigated the associations between these maternal factors and milk microbial composition and functionality. Methods: Women ≥20 years were recruited during pregnancy and milk samples were collected at 3 months post-partum (NCT01405547). Demographic data, weight, height, and a 3-hour oral glucose tolerance test were conducted at 30 (95% CI: 25-33) weeks gestation. Metagenomic DNA extraction and 16S ribosomal RNA gene sequencing of the V4 hypervariable region (Illumina MiSeq) was carried out on 113 milk samples. Results: Multivariable linear regression analyses demonstrated no significant associations between maternal characteristics (maternal BMI [pre-pregnancy, 3 months post-partum], glucose tolerance, mode of delivery and ethnicity) and microbiota alpha-diversity; however, pre-pregnancy BMI was associated with human milk beta-diversity (Bray-Curtis p=0.040). Women with a pre-pregnancy BMI >30 kg/m2 (obese) had a greater incidence of Bacteroidetes (incidence rate ratio [IRR]: 3.70 [95% CI: 1.61-8.48]) and a reduced incidence of Proteobacteria (0.62 [0.43-0.90]), compared to overweight women (BMI 25.0-29.9 kg/m2) as assessed by multivariable Poisson regression. Increased incidence of Gemella was observed among overweight (versus healthy) mothers with gestational diabetes (5.96 [1.85-19.21]) and obese (versus healthy) mothers with impaired glucose tolerance (4.04 [1.63-10.01]). An increased incidence of Brevundimonas (16.70 [5.99-46.57]) was found in the milk of women who underwent an unscheduled C-section versus vaginal delivery. Lastly, functional gene inference demonstrated that obesity was associated with increased abundance of genes encoding for the biosynthesis of secondary metabolites in milk (coefficient=0.00028, p=0.0070). Conclusions: Mother’s milk has a diverse microbiota of which its diversity and differential abundance appear associated with maternal body size, glucose tolerance status, mode of delivery, and ethnicity. Further research is warranted to determine whether this variability in the milk microbiota impacts colonization of the infant gut.