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

Abstract

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, we systematically investigated the associations between these maternal factors in women ≥20 years and milk microbial composition and predicted functionality by V4-16S ribosomal RNA gene sequencing (NCT01405547; https://clinicaltrials.gov/ct2/show/NCT01405547 ). Demographic data, weight, height, and a 3-h oral glucose tolerance test were gathered at 30 (95% CI: 25-33) weeks gestation, and milk samples were collected at 3 months post-partum (n = 113).

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 milk microbiota alpha-diversity; however, pre-pregnancy BMI was associated with human milk microbiota beta-diversity (Bray-Curtis R² = 0.037). 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]) in their milk, compared to women with an overweight BMI (25.0-29.9 kg/m²) as assessed by multivariable Poisson regression. An increased incidence of Gemella was observed among mothers with gestational diabetes who had an overweight BMI versus healthy range BMI (5.96 [1.85-19.21]). An increased incidence of Gemella was also observed among mothers with impaired glucose tolerance with an obese BMI versus mothers with a healthy range BMI (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 pre-pregnancy obesity was associated with an increased abundance of genes encoding for the biosynthesis of secondary metabolites pathway in milk (coefficient = 0.0024, P_FDR < 0.1).

Conclusions: Human milk has a diverse microbiota of which its diversity and differential abundance appear associated with maternal BMI, 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.

Keywords: Body mass index; Caesarean delivery; Ethnicity; Gestational diabetes; Human milk; Impaired glucose tolerance; Microbiome; Microbiota; Mode of delivery; Vaginal delivery.

Fig. 1

Microbial relative abundance in human milk at the phylum level (n = 109). The relative abundances of bacterial phyla in collected human milk samples are visualized using bar plots. For simplicity, only the most abundant 5 phyla are displayed with other phyla merged into the Other category

Fig. 2

Microbial relative abundance in human milk at the genus level (n = 109). The relative abundances of bacterial genera in collected human milk samples are visualized using bar plots. For simplicity, only the most abundant 10 genera are displayed with other genera merged into the Other category

Fig. 3

a-e The association between maternal characteristics and human milk microbiota alpha-diversity. The bacterial richness (Chao1 index) and diversity (Shannon index) of each human milk sample are plotted using box and whisker plots (mid-line = median; upper and lower bounds of the box = first and third quartile) as a function of a maternal glucose tolerance, b mode of delivery, c pre-pregnancy BMI, d 3-month post-partum BMI, e ethnicity. Multivariable linear regression analyses revealed no significant associations between the alpha-diversity of the milk microbiota and maternal metabolic and obstetrical characteristics. Abbreviations: GDM, gestational diabetes mellitus, IGT, impaired glucose tolerance; Sched CS, scheduled C-section; Unsched CS, unscheduled C-section