1H NMR-based metabolomics on human mature milk reveals the effects of oral VLS#3 probiotic supplementation to mothers

Background Perinatal manipulation of maternal gut microflora has been recently proposed as a possible dietary strategy to reduce the risk of disease in the infant. However, how a modulation of gut microbiota can affect the host physiology, the breast milk composition, and finally the infant physiology is still unclear. We applied a metabolomics approach, based on 1H NMR spectroscopy and multivariate analysis, to characterize the mature breast milk metabolome. Furthermore, we evaluated the feasibility of revealing the changes in breast milk composition induced by oral VLS#3 probiotic mixture supplementation. Methodology Pregnant women have been administered perinatally with either oral VLS#3 or placebo. Breast milk was drawn one month after delivery, extracted into polar and organic phases and analysed by 1H NMR spectroscopy. The metabolic profiles were defined by applying statistical multivariate methods on the collected spectra. Immunological assays were performed on the same milk samples to determine the interleukin levels. Principal findings 1H NMR spectra allowed to classify the milk samples on the basis of Lewis epitopes. VLS#3 supplementation to mothers induced a different milk metabolome characterized by a significant decrease in the levels of some fucosyl-oligosaccharides and citrate levels, and an increase in MUFA, phosphatidylglycerols, and succinate levels. Milks from probiotic supplemented mothers showed a significant increase in TGF- and IL-10 levels. Conclusions 1H-NMR-based global metabolic profiling is a powerful analytical approach to investigate the subtle changes on human milk induced by a diet intervention. Our results can have important implications to understand how the modulation of mother gut microbiota affects the breast milk composition, and thence the infant health.